{"product_id":"krasg12c-his-tag-protein-human-ua085047","title":"KRAS(G12C) 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%;\"\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\u003cp\u003eThr2-Cys185(G12C) 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%;\"\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\u003cp style=\"margin-bottom: 0px;\"\u003e20-25kDa (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＞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\u003cp\u003e50mM Tris, 200mM NaCl, 20% Glycerol, 1mM DTT, pH7.5\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\u003eStable for 12 months upon stored at -80℃ from the date of receipt. And avoid repeated freeze-thaws 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.Kim D, Xue JY, Lito P. Targeting KRAS(G12C): From Inhibitory Mechanism to Modulation of Antitumor Effects in Patients. Cell. 2020 Nov 12;183(4):850-859.\u003cbr\u003e2.Canon J, Rex K, Saiki AY, Mohr C, Cooke K, Bagal D, Gaida K, Holt T, Knutson CG, Koppada N, Lanman BA, Werner J, Rapaport AS, San Miguel T, Ortiz R, Osgood T, Sun JR, Zhu X, McCarter JD, Volak LP, Houk BE, Fakih MG, O'Neil BH, Price TJ, Falchook GS, Desai J, Kuo J, Govindan R, Hong DS, Ouyang W, Henary H, Arvedson T, Cee VJ, Lipford JR. The clinical KRAS(G12C) inhibitor AMG 510 drives anti-tumour immunity. Nature. 2019 Nov;575(7781):217-223.\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\u003eKRAS(G12C) is an oncogenic mutant of the RAS family member KRAS, in which a glycine at position 12 is mutated to cysteine. This mutation disrupts GTP hydrolysis, causing the protein to remain persistently in the GTP-bound \"activated state\". The G12C mutation introduces a nucleophilic cysteine into the Switch II pocket, providing a binding site for covalent inhibitors (e.g., Sotorasib, Adagrasib), which lock KRAS in the inactive GDP-bound conformation. This mutation constitutively activates downstream pathways such as RAF-MEK-ERK and PI3K-AKT, driving cell proliferation and survival, making it a key driver in non-small cell lung cancer (NSCLC; ~14%), pancreatic cancer (~2%), and colorectal cancer (~3%). Clinically, KRAS(G12C) was once considered an \"undruggable\" target until the first covalent inhibitor, AMG 510, emerged in 2019, marking a major breakthrough in precision oncology. To date, the FDA has approved Sotorasib and Adagrasib for second-line treatment of KRAS G12C-mutant NSCLC. Combination therapy with EGFR inhibitors has also shown significant efficacy in colorectal cancer; however, acquired resistance remains a clinical challenge.\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 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\u003cstrong\u003eMaterials\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e1.\u003c\/span\u003e\u003cspan\u003eKRAS(G12C) 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\u003cstrong\u003eProduce\u003c\/strong\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\u003ctable style=\"border-collapse:collapse;border:1px solid #000000;width:auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"border:1px solid #000000;text-align:center;vertical-align:middle;\"\u003e\u003cp style=\"text-align:center;\"\u003e\u003cstrong\u003eSample Name\u003c\/strong\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\u003cstrong\u003eAmount\u003c\/strong\u003e\u003c\/p\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\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eGTPase Glo ™ Reagent, 500X\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\u003e2 μL\u003c\/span\u003e\u003c\/p\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\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eADP, 10 mM\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\u003e0.5 μL\u003c\/span\u003e\u003c\/p\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\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eGTPase Glo ™ Buffer\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\u003e998 μL\u003c\/span\u003e\u003c\/p\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\u003cp style=\"text-align:center;\"\u003e\u003cspan\u003eTotal volume\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 style=\"margin-right:8px;\"\u003e1\u003c\/span\u003e\u003cspan\u003emL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/figure\u003e\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\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\u003e(1-Sample OD\/BLANK OD)*50pmol\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\u003eIncubation time(min) ×amount 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\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 style=\"text-align:left;\"\u003e \u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"10μg","offer_id":43093483716683,"sku":"UA085047-10μg","price":250.0,"currency_code":"USD","in_stock":true},{"title":"50μg","offer_id":43093483749451,"sku":"UA085047-50μg","price":500.0,"currency_code":"USD","in_stock":true},{"title":"100μg","offer_id":43093483782219,"sku":"UA085047-100μg","price":800.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/fd6fb1c871dd46efa3091393287be66c.png?v=1788192223","url":"https:\/\/www.antbioinc.com\/products\/krasg12c-his-tag-protein-human-ua085047","provider":"AntBio","version":"1.0","type":"link"}