{"product_id":"idh1-his-tag-protein-human-ua085026","title":"IDH1 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%;\"\u003eIDH1, PICD, IDP\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%;\"\u003eO75874-1\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\u003eMet 1 - Leu 414 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;\"\u003e40-55kDa (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,\u0026gt; 90% 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%;\"\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, 150mM NaCl, pH7.5, 1mM DTT, 10%Glycerol\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\n\u003cp\u003eStable for 12 months upon stored at -80℃ from the date of receipt. \u003c\/p\u003e\n\u003cp\u003eAnd avoid repeated freeze-thaws cycles.\u003cbr\u003e\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\u003cp\u003e1. Parsons, D.W., et al. (2008). An integrated genomic analysis of human glioblastoma multiforme. Science, 321(5897), 1807-1812.\u003cbr\u003e2. Dang, L., et al. (2009). Cancer-associated IDH1 mutations produce 2-hydroxyglutarate. Nature, 462(7274), 739-744.\u003cbr\u003e3. Xu, W., et al. (2011). Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of α-ketoglutarate-dependent dioxygenases. Cancer Cell, 19(1), 17-30.\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\u003cspan\u003e\u003cspan\u003eIDH1 (Isocitrate Dehydrogenase 1) is a key metabolic enzyme located in the cytoplasm and peroxisomes. Its primary physiological function is to catalyze the oxidative decarboxylation of isocitrate to produce alpha-ketoglutarate (α-KG), while concurrently reducing NADP+ to NADPH. This reaction is crucial for cellular metabolism, as it links the tricarboxylic acid (TCA) cycle with lipid metabolism and provides NADPH, a major reducing agent essential for antioxidant defense and anabolic biosynthesis.\u003c\/span\u003e\u003cspan\u003eA seminal discovery in cancer biology revealed that specific heterozygous somatic mutations in the IDH1 gene, most commonly at the R132 residue, are frequent drivers in several cancers. These include gliomas (especially secondary glioblastoma and oligodendroglioma), acute myeloid leukemia (AML), cholangiocarcinoma, and chondrosarcomas. The mutant enzyme acquires a neomorphic activity: instead of producing α-KG, it reduces α-KG to D-2-hydroxyglutarate (D-2HG). This oncometabolite, D-2HG, accumulates to high levels and functions as a competitive inhibitor of multiple α-KG-dependent dioxygenases. This inhibition leads to profound epigenetic dysregulation (via inhibition of histone and DNA demethylases) and blockade of cellular differentiation, thereby promoting tumorigenesis.\u003c\/span\u003e\u003c\/span\u003e\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003eAssay protocol\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003ePrinciple: Measured by the ability to oxidatively decarboxylate isocitrate to 2-oxoglutarate.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003eMaterials\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e1.     Assay Buffer\u003c\/span\u003e\u003cspan\u003e：\u003c\/span\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e25 mM Tris, 0.5 mM MnCl2, 5 mM DTT, pH 7.5\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e2.     IDH1 His Tag Protein, Human\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e3.     Substrate1: DL-Isocitricacid (MCE, Catalog # HY-W009362)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e4.     Substrate2: NADP+ (Aladdin, Catalog # N101669) \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e5.     96 Well Clear Plate (BIOFIL, Catalog#011096)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e6.     Plate Reader (PerkinElmer, ABS,340 nm, kinetic mode,60s\/cycle,10 cycles)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003eProduce\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e1. Dilute IDH1 to 0.8 μg\/mL, 0.4 μg\/mL in Assay Buffer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e2. Prepare a Substrate Mixture by Diluting NADP+ and Isocitric Acid to 1 mM and 2 mM, respectively, in Assay Buffer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e3. Load into a plate 50 μL of dilute IDH1 protein and start the reaction by adding 50 μL of Substrate Mix. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of Substrate Mix.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e4. Read plate at a wavelength of 340 nm (bottom read) in kinetic mode for 10 minutes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e5. Calculate specific activity.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003eSpecific Activity (pmol\/min\/µg) =   Slope (OD\/min) x well volume (L) x 1012pmol\/mol\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003e        ext. coeff (M-1cm-1) x path corr. (cm) x amount of enzyme (μg)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003eSlope\u003c\/span\u003e\u003cspan\u003e：\u003c\/span\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003eAdjusted for Substrate Blank\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003eext. coeff\u003c\/span\u003e\u003cspan\u003e：\u003c\/span\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003eUsing the extinction coefficient 6270 M-1cm-1\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003epath corr\u003c\/span\u003e\u003cspan\u003e：\u003c\/span\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;' lang=\"EN-US\"\u003eUsing the path correction 0.320 cm\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"10μg","offer_id":42332765028427,"sku":null,"price":100.0,"currency_code":"USD","in_stock":true},{"title":"50μg","offer_id":42332765061195,"sku":null,"price":310.0,"currency_code":"USD","in_stock":true},{"title":"100μg","offer_id":42332765093963,"sku":null,"price":495.0,"currency_code":"USD","in_stock":true},{"title":"500μg","offer_id":42332765126731,"sku":null,"price":1800.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_9b1a38cf-28e6-41fb-950e-e2a5e4f931b3.png?v=1776073666","url":"https:\/\/www.antbioinc.com\/products\/idh1-his-tag-protein-human-ua085026","provider":"AntBio","version":"1.0","type":"link"}