{"product_id":"krasg12d-his-tag-protein-human-ua085040","title":"KRAS(G12D) His Tag Protein, Human","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\n\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\n\u003ctable style=\"width: 100%; height: auto;\"\u003e\n\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%;\"\u003eK-Ras 2, Ki-Ras, c-K-ras, c-Ki-ras, GTPase KRas, KRAS2, RASK2\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%;\"\u003eP01116-2\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\n\u003cp\u003eThr2-Cys185（G12D）with His Tag at the C-Terminus\u003c\/p\u003e\n\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%;\"\u003eE.coli\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\n\u003cp style=\"margin-bottom: 0px;\"\u003e20-25kDa (Reducing)\u003c\/p\u003e\n\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＞95% by SDS-PAGE\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%;\"\u003eLiquid\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\n\u003cp\u003e20 mM Tris, 150 mM NaCl, 1 mM DTT, PH7.4 ,10% glycerol\u003c\/p\u003e\n\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\n\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\n\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\n\u003cp\u003eStable for 12 months upon stored at -80℃ from the date of receipt. And avoid repeated freeze-thaws cycles.\u003c\/p\u003e\n\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\n\u003cp\u003e1. Gremer L, Merbitz-Zahradnik T, Dvorsky R, Cirstea IC, Kratz CP, Zenker M, Wittinghofer A, Ahmadian MR. Germline KRAS mutations cause aberrant biochemical and physical properties leading to developmental disorders. Hum Mutat. 2011 Jan;32(1):33-43.\u003cbr\u003e2. Serra RW, Fang M, Park SM, Hutchinson L, Green MR. A KRAS-directed transcriptional silencing pathway that mediates the CpG island methylator phenotype. Elife. 2014 Mar 12;3:e02313. \u003cbr\u003e3. Sun Q, Burke JP, Phan J, Burns MC, Olejniczak ET, Waterson AG, Lee T, Rossanese OW, Fesik SW. Discovery of small molecules that bind to K-Ras and inhibit Sos-mediated activation. Angew Chem Int Ed Engl. 2012 Jun 18;51(25):6140-3. \u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cdiv\u003e\n\u003cp\u003e\u003cspan\u003eThe KRAS protein (Kirsten rat sarcoma viral oncogene homolog) is a small GTPase protein whose structure comprises a G-domain (responsible for GTP\/GDP binding) and a hypervariable region. The G12D mutation refers to the substitution of glycine at position 12 (Gly12) with aspartic acid (Asp), which prevents GAP (GTPase-activating protein) from promoting GTP hydrolysis, thereby locking KRAS in a persistently GTP-bound \"active\" state. This leads to constitutive activation of downstream signaling pathways including RAF-MEK-ERK and PI3K-AKT, driving cellular proliferation, survival, and metabolic reprogramming. Clinically, KRAS G12D represents one of the most prevalent oncogenic mutations in pancreatic cancer (~40%), colorectal cancer, and lung cancer, and has historically been considered \"undruggable.\" However, recent breakthroughs include the development of non-covalent inhibitors (binding the Switch II pocket via salt bridge formation), elucidation of synergistic tumorigenic mechanisms between PTEN loss and G12D, demonstration of G12D-induced immunosuppressive microenvironment conferring resistance to PD-1\/PD-L1 inhibitors, and discovery of G12D-specific stem cell reprogramming in lung adenocarcinoma, offering novel directions for precision-targeted therapies and combination immunotherapeutic strategies.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n\u003ch4\u003eProtocol\u003c\/h4\u003e\n\u003cdiv\u003e\n\u003cp style=\"text-align: left;\"\u003e\u003cstrong\u003eAssay protocol\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003ePrinciple: The GTPase Glo™ Assay assesses the activities of KRAS by detecting the amount of GTP remaining after GTP hydrolysis in a KRAS reaction.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eMaterials\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003eKRAS(G12D) His Tag Protein, Human\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003e\u003cspan\u003eGTPase Glo™ Assay (Promega, Catalog # V7681T)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\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;\"\u003e4.\u003c\/span\u003e\u003cspan\u003ePlate Reader (PerkinElmer)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eProduce\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003ePrepare a 2X GTP solution containing 10 µM GTP and 2mM DTT in GTPase\/GAP Buffer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003e\u003cspan\u003eDilute the KRAS to 100 µg\/mL、80 µg\/mL and 60 µg\/mL in GTPase\/GAP Buffer and dispense 5 µL into each well of a 384 well plate.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\u003c\/span\u003e\u003cspan\u003eInitiate the reaction by adding 5 µL of the 2X GTP solution prepared in Step 1 to each well. Include a 2X GTP solution with 5 µL GTPase\/GAP Buffer as Blank. The reaction volume is 10 µL.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4.\u003c\/span\u003e\u003cspan\u003eIncubate the reaction at room temperature (22-25℃) for 30 minutes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5.\u003c\/span\u003e\u003cspan\u003eGently mix the thawed GTPase Glo™ Reagent, 500X, by inversion; do not vortex. Prepare the required volume of reconstituted GTPase Glo™ Reagent by increasing or decreasing the component volumes provided below.\u003c\/span\u003e\u003c\/p\u003e\n\u003cfigure class=\"table\"\u003e\n\u003ctable style=\"border-collapse: collapse; border: 1px solid #000000; width: auto;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003eSample Name\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003eAmount\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003eGTPase Glo ™ Reagent, 500X\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003e2 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003eADP, 10 mM\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003e0.5 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003eGTPase Glo ™ Buffer\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003e998 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003eTotal volume\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1\u003c\/span\u003e\u003cspan\u003emL\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/figure\u003e\n\u003cp\u003e\u003cbr\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6.\u003c\/span\u003e\u003cspan\u003eAdd 10 µL of reconstituted GTPase-Glo™ Reagent to the completed reaction, mix briefly and incubate with shaking for 30 minutes at room temperature (22–25℃).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e7.\u003c\/span\u003e\u003cspan\u003eAdd 20 µL of Detection Reagent and incubate the plate for 5-10 minutes at room temperature (22–25℃).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e8.\u003c\/span\u003e\u003cspan\u003eRead at emission wavelengths of 555 nm (luminescence), respectively in endpoint mode.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e9.\u003c\/span\u003e\u003cspan\u003eCalculate specific activity.\u003c\/span\u003e\u003c\/p\u003e\n\u003cfigure class=\"table\"\u003e\n\u003ctable style=\"border-collapse: collapse; border: 1px solid #000000; width: auto;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\" rowspan=\"2\"\u003e\u003cspan\u003eSpecific Activity (pmol\/min\/μg) =\u003c\/span\u003e\u003c\/td\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\u003cspan\u003e(1-Sample OD\/BLANK OD)*50pmol\u003c\/span\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"border: 1px solid #000000; text-align: center; vertical-align: middle;\"\u003e\u003cspan\u003eIncubation time(min) ×amount of enzyme (μg)\u003c\/span\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/figure\u003e\n\u003cp\u003e\u003cbr\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eSample OD: Remains of ATP OD\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eBLANK OD: Added of GTP OD\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e50pmol: Added of GTP\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eIncubation time: 30 min\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eamount of enzyme: 0.5μg, 0.4 μg and 0.3 μg\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental method\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eExperimental Principle: The GTPase Glo™ Assay evaluates KRAS activity by measuring the amount of remaining GTP after GTP hydrolysis in the KRAS reaction.\u003cbr\u003e\u003cstrong\u003eExperimental Materials\u003c\/strong\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003eKRAS(G12D) His Tag Protein, Human\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003e\u003cspan\u003eGTPase Glo™ Assay (Promega, Catalog # V7681T)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\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;\"\u003e4.\u003c\/span\u003e\u003cspan\u003ePlate Reader (PerkinElmer)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"10μg","offer_id":42332771483723,"sku":null,"price":195.0,"currency_code":"USD","in_stock":true},{"title":"50μg","offer_id":42332771516491,"sku":null,"price":390.0,"currency_code":"USD","in_stock":true},{"title":"100μg","offer_id":42332771549259,"sku":null,"price":700.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_f625358b-c26e-40e7-b35b-ba74f43de101.png?v=1776073834","url":"https:\/\/www.antbioinc.com\/products\/krasg12d-his-tag-protein-human-ua085040","provider":"AntBio","version":"1.0","type":"link"}