{"product_id":"irak4-his-tag-protein-human-ua085033","title":"IRAK4 His Tag Protein, Human","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSpecies\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eHuman\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eIRAK-4,IMD67,IPD1,NY-REN-64,REN64\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eAccession\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eQ9NWZ3\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eAmino Acid Sequence\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eGlu154-Ser460 with His Tag at the C-Terminus\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eExpression System\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eBaculovirus-InsectCells\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eMolecular Weight\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e33-43kDa (Reducing)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePurity\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e＞90% by SDS-PAGE,\u0026gt; 85% by HPLC.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eConjugation\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eUnconjugated\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eTag\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eHis Tag\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePhysical Appearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eLyophilized powder\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStorage Buffer\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003ePBS, PH7.4, 5% trehalose\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReconstitution\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eReconstitute at 0.1-1 mg\/ml according to the size in ultrapure water after rapid centrifugation.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e· 12 months from date of receipt, lyophilized powder stored at -20 to -80℃.\u003cbr\u003e· 3 months, -20 to -80℃ under sterile conditions after reconstitution.\u003cbr\u003e· 1 week, 2 to 8℃ under sterile conditions after reconstitution.\u003cbr\u003e· Please avoid repeated freeze-thaw cycles.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReference\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e1.Wright HJ, Hou J, Xu B, Cortez M, Potma EO, Tromberg BJ, Razorenova OV. CDCP1 drives triple-negative breast cancer metastasis through reduction of lipid-droplet abundance and stimulation of fatty acid oxidation. Proc Natl Acad Sci U S A. 2017 Aug 8;114(32):E6556-E6565.\u003cbr\u003e2.Ackerman L, Acloque G, Bacchelli S, Schwartz H, Feinstein BJ, La Stella P, Alavi A, Gollerkeri A, Davis J, Campbell V, McDonald A, Agarwal S, Karnik R, Shi K, Mishkin A, Culbertson J, Klaus C, Enerson B, Massa V, Kuhn E, Sharma K, Keaney E, Barnes R, Chen D, Zheng X, Rong H, Sabesan V, Ho C, Mainolfi N, Slavin A, Gollob JA. IRAK4 degrader in hidradenitis suppurativa and atopic dermatitis: a phase 1 trial. Nat Med. 2023 Dec;29(12):3127-3136.\u003cbr\u003e3.Kawagoe T, Sato S, Jung A, Yamamoto M, Matsui K, Kato H, Uematsu S, Takeuchi O, Akira S. Essential role of IRAK-4 protein and its kinase activity in Toll-like receptor-mediated immune responses but not in TCR signaling. J Exp Med. 2007 May 14;204(5):1013-24.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cspan\u003eIRAK4 (Interleukin-1 Receptor-Associated Kinase 4) is a serine\/threonine kinase. Its N-terminal death domain facilitates assembly with MyD88 and IRAK2 into a 6:4:4 helical Myddosome signaling complex, while its C-terminal kinase domain is activated through dimerization and autophosphorylation. A unique tyrosine gatekeeper residue provides a strategic advantage for the development of selective inhibitors. As a central component of the TLR\/IL-1R signaling pathway, IRAK4 activates NF-κB and MAPK pathways to regulate innate immune responses. Deficiency in IRAK4 leads to specific susceptibility to pyogenic bacteria (such as\u003c\/span\u003e\u003cspan\u003e Streptococcus pneumoniae) while preserving normal resistance to viral and fungal infections. Clinically, IRAK4 is not only implicated as a causative gene in primary immunodeficiency disorders but also plays a critical role in driving tumor survival in lymphomas harboring MYD88 mutations. Currently, IRAK4-targeted small-molecule inhibitors and PROTAC degraders have advanced into clinical trials for the treatment of rheumatoid arthritis, atopic dermatitis, hidradenitis suppurativa, and B-cell lymphomas. These therapeutic agents achieve precise immunomodulation by either blocking kinase activity or eliminating scaffold functions.\u003c\/span\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003eAssay protocol\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003ePrinciple:\u003c\/span\u003e\u003cspan\u003e The IRAK4 assay is performed using the ADP-Glo\u003c\/span\u003e\u003cspan\u003eTM\u003c\/span\u003e\u003cspan\u003e Kinase Assay kit which quantifies the amount of ADP produced by the IRAK4 reaction. The ADP-Glo\u003c\/span\u003e\u003cspan\u003eTM \u003c\/span\u003e\u003cspan\u003eReagent is added to terminate the kinase reaction and to deplete the remaining ATP, and then the Kinase Detection Reagent is added to convert ADP to ATP and to measure the newly synthesized ATP using luciferase\/luciferin reaction.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003eMaterials\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003eKinase assay buffer(5X): 200 mM Tris-HCl, pH 7.4, 100 mM MgCl\u003c\/span\u003e\u003cspan\u003e2\u003c\/span\u003e\u003cspan\u003e and 0.5 mg\/mL BSA, 250 μM DTT\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003e\u003cspan\u003eKinase assay buffer(1X): 40 mM Tris-HCl, pH 7.4, 20 mM MgCl\u003c\/span\u003e\u003cspan\u003e2\u003c\/span\u003e\u003cspan\u003e, 0.1 mg\/mL BSA, 50 μM DTT\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\u003c\/span\u003e\u003cspan\u003eIRAK4 His Tag Protein, Human\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4.\u003c\/span\u003e\u003cspan\u003eADP-Glo Kinase Assay (Promega, Catalog # V6930)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5.\u003c\/span\u003e\u003cspan\u003eSubstrate: Myelin Basic Protein (MBP) (Sinobiological, Catalog # M42-51N)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6.\u003c\/span\u003e\u003cspan\u003eSolid white multi-well plate (384-well plate) (Corning, Catalog #3572)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e7.\u003c\/span\u003e\u003cspan\u003ePlate Reader (PerkinElmer)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003eProduce\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003ePrepare a substrate\/ATP mixture as follows (25 μM example).\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eSample Name\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eAmount (μL)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e10 mM ATP Solution\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eKinase Assay Buffer III (5x)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e79\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eSubstrate at 0.5 mg\/mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e80\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\u003cp\u003e\u003cspan\u003e2. Dilute the\u003c\/span\u003e \u003cspan\u003eIRAK4 to 40 µg\/mL, 20 µg\/mL and 10 µg\/mL in Kinase Assay Buffer (1x) and dispense 3 μL into each well of a 384-well plate.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e3. Initiate the reaction by adding 2 μL of the detection system prepared in Step 1 to each well. Include a detection system with 3 μL Kinase Assay Buffer (1x) as Blank. The reaction volume is 5 μL.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e4. Incubate the reaction at room temperature (22–25℃) for 40 minutes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e5.Add 5 μL of ADP-Glo Reagent to the completed reaction, mix briefly and incubate for 40 minutes at room temperature (22–25℃).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e6.Add 10 μL of Detection Reagent and incubate the plate for 30 minutes at room temperature (22–25℃).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e7.Read at luminescence, respectively in endpoint mode.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e8.Calculate specific activity.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003e• Standard Curve\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eDilute the ATP and ADP to 25 μM in Kinase Assay Buffer (1x).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e1.\u003c\/span\u003e\u003cspan\u003eMix 25 μΜ ATP and 25 μM ADP to form an ATP+ADP solution provided below and dispense 5 μL into each well of a 384-well plate.\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eWell Number\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e3\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e4\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e5\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e6\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e7\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e8\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e9\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e10\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e11\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e12\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e25μM ADP (μL)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e100\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e80\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e60\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e40\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e20\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e10\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e5\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e4\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e3\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e25μM ATP (μL)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e20\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e40\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e60\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e80\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e90\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e95\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e96\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e97\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e98\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e99\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e100\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\u003cp\u003e\u003cspan\u003e2.\u003c\/span\u003e\u003cspan\u003eAdd 5 μL of ADP-Glo Reagent to the completed reaction, mix briefly and incubate for 40 minutes at room temperature (22–25℃).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e3.\u003c\/span\u003e\u003cspan\u003eAdd 10 μL of Detection Reagent and incubate the plate for 30 minutes at room temperature (22–25℃).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e4.\u003c\/span\u003e\u003cspan\u003eRead at luminescence, respectively in endpoint mode.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e5.\u003c\/span\u003e\u003cspan\u003eDetect optical signals and establish conversion curves.\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse:collapse;width:auto;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eSpecific Activity (pmol\/min\/μg) =\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eATP (pmol)-Blank\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIncubation time(min) ×amount of enzyme (μg)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\u003cp\u003e\u003c\/p\u003e\n\u003cp\u003e\u003c\/p\u003e\n\u003cp\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003eExperimental Method\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003eExperimental Principle\u003c\/span\u003e\u003cspan\u003e: The IRAK4 activity assay is performed using the ADP-Glo™ Kinase Assay kit, which quantifies the amount of ADP produced by the IRAK4 reaction. The specific steps are as follows: First, add ADP-Glo™ Reagent to terminate the kinase reaction and deplete remaining ATP; then add the Kinase Detection Reagent to convert ADP to ATP, and measure the newly synthesized ATP using the luciferase\/luciferin reaction system.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003eExperimental Materials\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e1.\u003c\/span\u003e\u003cspan\u003eKinase assay buffer(5X): 200 mM Tris-HCl, pH 7.4, 100 mM MgCl\u003c\/span\u003e\u003cspan\u003e2\u003c\/span\u003e\u003cspan\u003e and 0.5 mg\/mL BSA, 250 μM DTT\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e2.\u003c\/span\u003e\u003cspan\u003eKinase assay buffer(1X): 40 mM Tris-HCl, pH 7.4, 20 mM MgCl\u003c\/span\u003e\u003cspan\u003e2\u003c\/span\u003e\u003cspan\u003e, 0.1 mg\/mL BSA, 50 μM DTT\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e3.\u003c\/span\u003e\u003cspan\u003eIRAK4 His Tag Protein, Human\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e4.\u003c\/span\u003e\u003cspan\u003eADP-Glo Kinase Assay (Promega, Catalog # V6930)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e5.\u003c\/span\u003e\u003cspan\u003eSubstrate: Myelin Basic Protein (MBP) (Sinobiological, Catalog # M42-51N)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e6.\u003c\/span\u003e\u003cspan\u003eSolid white multi-well plate (384-well plate) (Corning, Catalog #3572)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e7.\u003c\/span\u003e\u003cspan\u003ePlate Reader (PerkinElmer)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003eExperimental Steps\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e1.\u003c\/span\u003e\u003cspan\u003ePrepare substrate\/ATP mixture as follows (example: 25 μM):\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eSample Name\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eAmount (μL)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e10 mM ATP Solution\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eKinase Assay Buffer III (5x)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e79\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eSubstrate at 0.5 mg\/mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e80\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\u003cp\u003e\u003cspan\u003e2.\u003c\/span\u003e\u003cspan\u003eDilute IRAK4 to 40 µg\/mL, 20 µg\/mL, and 10 µg\/mL in Kinase Assay Buffer (1x) and dispense 3 μL into each well of a 384-well plate.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e3.\u003c\/span\u003e\u003cspan\u003eInitiate the reaction by adding\u003c\/span\u003e\u003cspan\u003e2\u003c\/span\u003e\u003cspan\u003e μL of the detection system prepared in Step 1 to each well. Include a detection system with 3 μL Kinase Assay Buffer (1x) as Blank. The reaction volume is 5 μL. Incubate the reaction at room temperature (22–25°C) for 40 minutes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e4.\u003c\/span\u003e\u003cspan\u003eAdd 5 μL of ADP-Glo Reagent to the completed reaction, mix briefly and incubate for 40 minutes at room temperature (22–25°C).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e5.\u003c\/span\u003e\u003cspan\u003eAdd 10 μL Detection Reagent and incubate the plate for 30 minutes at room temperature (22–25°C).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e6.\u003c\/span\u003e\u003cspan\u003eRead chemiluminescence signals respectively in endpoint mode.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e7.\u003c\/span\u003e\u003cspan\u003eCalculate specific activity.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003eStandard Curve\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e1.\u003c\/span\u003e\u003cspan\u003eDilute ATP and ADP to 25 μM in Kinase Assay Buffer (1x).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e2.\u003c\/span\u003e\u003cspan\u003eMix 25 μM ATP and 25 μM ADP according to the table below to form an ATP+ADP solution, and dispense 5 μL into each well of a 384-well plate.\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eWell Number\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e3\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e4\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e5\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e6\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e7\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e8\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e9\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e10\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e11\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e12\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e25μM ADP (μL)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e100\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e80\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e60\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e40\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e20\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e10\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e5\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e4\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e3\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e25μM ATP (μL)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e20\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e40\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e60\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e80\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e90\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e95\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e96\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e97\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e98\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e99\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e100\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\u003cp\u003e\u003cspan\u003e3.\u003c\/span\u003e\u003cspan\u003eAdd 5 μL of ADP-Glo Reagent to the completed wells, mix briefly and incubate for 40 minutes at room temperature (22-25°C).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e4.\u003c\/span\u003e\u003cspan\u003eAdd 10 μL Detection Reagent and incubate the plate for 30 minutes at room temperature (22-25°C).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e5.\u003c\/span\u003e\u003cspan\u003eRead chemiluminescence signals respectively in endpoint mode.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e6.\u003c\/span\u003e\u003cspan\u003eDetect optical signals and establish conversion curves.\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse:collapse;width:auto;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eSpecific Activity (pmol\/min\/μg) =\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eATP (pmol)-Blank\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIncubation time(min) ×amount of enzyme (μg)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\u003cp\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"10μg","offer_id":42332766568523,"sku":null,"price":185.0,"currency_code":"USD","in_stock":true},{"title":"25μg","offer_id":42332766601291,"sku":null,"price":435.0,"currency_code":"USD","in_stock":true},{"title":"50μg","offer_id":42332766634059,"sku":null,"price":695.0,"currency_code":"USD","in_stock":true},{"title":"100μg","offer_id":42332766666827,"sku":null,"price":1110.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_9ee3bdfa-0390-4919-9e2e-9d12b68f44f2.png?v=1780633781","url":"https:\/\/www.antbioinc.com\/products\/irak4-his-tag-protein-human-ua085033","provider":"AntBio","version":"1.0","type":"link"}