{"product_id":"mmp-9-his-tag-protein-mouse-ua016087","title":"MMP-9 His Tag Protein, Mouse","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%;\"\u003eMouse\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%;\"\u003eMMP9, CLG4B, GELB, MANDP2, Gelatinase B\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%;\"\u003eP41245\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\u003eAla20-Pro730 with His Tag at 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%;\"\u003eHEK293\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%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003e100-130kDa (Reducing)\u003c\/p\u003e\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\u0026gt; 95% by SDS-PAGE,\u0026gt;90% by HPLC\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eEndotoxin\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u0026lt;0.1EU\/μg\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%;\"\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eReconstitute at 0.1-1 mg\/ml according to the size in ultrapure water after rapid centrifugation.\u003c\/p\u003e\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.Ji Y, Gao Q, Ma Y, Wang F, Tan X, Song D, Hoo RLC, Wang Z, Ge X, Han H, Guo F, Chang J. An MMP-9 exclusive neutralizing antibody attenuates blood-brain barrier breakdown in mice with stroke and reduces stroke patient-derived MMP-9 activity. Pharmacol Res. 2023 Apr;190:106720. \u003cbr\u003e2.Gough PJ, Gomez IG, Wille PT, Raines EW. Macrophage expression of active MMP-9 induces acute plaque disruption in apoE-deficient mice. J Clin Invest. 2006 Jan;116(1):59-69. \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\u003eMatrix metalloproteinase 9 (MMP-9), also known as 92 kDa type IV collagenase, 92 kDa gelatinase, gelatinase B (GELB), or CLG4B, is secreted by neutrophils, macrophages, and various transformed cells. It is the most complex member of the MMP family in terms of domain structure and activity regulation.Structurally, MMP-9 can be divided into five distinct domains: a prodomain that is cleaved upon activation, a gelatin-binding domain consisting of three contiguous fibronectin type II units, a catalytic domain with the zinc-binding site, a proline-rich linker region, and a carboxyl-terminal hemopexin-like domain. This enzyme degrades various substrates, including gelatin, collagen types IV and V, and elastin.MMP-9 is implicated in a variety of autoimmune diseases, such as systemic lupus erythematosus, rheumatoid arthritis, and multiple sclerosis, and is considered a potential therapeutic target.\u003c\/span\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cstrong\u003eAssay protocol\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003ePrinciple:\u003c\/strong\u003e\u003cspan\u003e Measured by its ability to cleave a peptide substrate, Mca-PLGL-Dpa-AR-NH2.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eMaterials\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003eAssay Buffer: 50 mM Tris, 10 mM CaCl\u003csub\u003e2\u003c\/sub\u003e, 150 mM NaCl, 0.05% (w\/v) Brij 35, pH 7.5 (TCNB)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e2.\u003c\/span\u003e\u003cspan\u003eMMP-9 His Tag Protein, Mouse\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e3.\u003c\/span\u003e\u003cspan\u003ep-aminophenylmercuric acetate (APMA) (Aladdin, Catalog # P649286)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e4.\u003c\/span\u003e\u003cspan\u003eFluorogenic Peptide Substrate: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (R\u0026amp;D, ES001)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e5.\u003c\/span\u003e\u003cspan\u003eCalibration standard: MCA-Pro-Leu-OH (Shyuanye, T77046)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e6.\u003c\/span\u003e\u003cspan\u003e96 ELISA Removable Plate, Black, High binding (GENEVER, Catalog # GMO2-96H)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e7.\u003c\/span\u003e\u003cspan\u003ePlate Reader (PerkinElmer, excitation 320 nm, and emission 405 nm)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eProduce\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan style=\"margin-right:8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003eActivate MMP-9 at 100 μg\/mL with 1 mM APMA in Assay Buffer. Incubate at 37 ℃ for 1 hour.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan style=\"margin-right:8px;\"\u003e2.\u003c\/span\u003e\u003cspan\u003eDilute activated MMP-9 (From the step 2) to 0.8, 0.4 and 0.2 μg\/mL in Assay Buffer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan style=\"margin-right:8px;\"\u003e3.\u003c\/span\u003e\u003cspan\u003eDilute Substrate to 20 μM in Assay Buffer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan style=\"margin-right:8px;\"\u003e4.\u003c\/span\u003e\u003cspan\u003eLoad 50 μL of the dilute MMP-9 Protein (From the step 2) into a black well plate and start the reaction by adding 50 μL of 20 μM Substrate. Include a Substrate with Assay Buffer as Blank.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan style=\"margin-right:8px;\"\u003e5.\u003c\/span\u003e\u003cspan\u003eRead at excitation and emission wavelengths of 320 nm and 405 nm, kinetic mode, 60s\/cycle, 20 cycles.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan style=\"margin-right:8px;\"\u003e6.\u003c\/span\u003e\u003cspan\u003eCalculate specific activity.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e• Standard Curve\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e1. Dilute Calibration standard to 10 μM in Assay Buffer and prepare serial dilutions.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e2. Add 100 μL of each serially diluted standard and blank (Assay buffer) into appropriate wells of a 96-well, the standard curve has a range of 1000, 500, 250, 125, 62.5, 31.25, 15.625 pmol per well.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e3. Read at excitation and emission wavelengths of 320 nm and 405 nm, respectively in endpoint mode.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e4. Linear Regression of MCA-Pro-Leu-OH (pmol)(y) – RFU-Blank(x).\u003c\/span\u003e\u003c\/p\u003e\n\u003cfigure class=\"table\"\u003e\u003ctable style=\"border-collapse:collapse;border:1px solid #000000;width:auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;\" rowspan=\"2\"\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=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eAdjusted Vmax (RFU\/min) x Conversion Factor (pmol\/RFU)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eamount of enzyme (µg)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/figure\u003e\u003cp\u003e\u003cbr\u003e \u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e \u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"25μg","offer_id":43094139535435,"sku":"UA016087-25μg","price":270.0,"currency_code":"USD","in_stock":true},{"title":"100μg","offer_id":43094139568203,"sku":"UA016087-100μg","price":550.0,"currency_code":"USD","in_stock":true},{"title":"500μg","offer_id":43094139600971,"sku":"UA016087-500μg","price":1070.0,"currency_code":"USD","in_stock":true},{"title":"1mg","offer_id":43094139633739,"sku":"UA016087-1mg","price":1600.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/862cedfbc9d64935a02765b465b21009.png?v=1788202833","url":"https:\/\/www.antbioinc.com\/products\/mmp-9-his-tag-protein-mouse-ua016087","provider":"AntBio","version":"1.0","type":"link"}