{"product_id":"mat1a-his-tag-protein-human-ua085073","title":"MAT1A 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%;\"\u003eAMS1,METK1\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%;\"\u003eQ00266\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\u003eMet1-Phe395 with His\u0026amp;Thrombin Tag at the N-Terminus\u003cbr\u003e\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\u003e40-55kDa (Reducing)\u003cbr\u003e\u003c\/p\u003e\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;1EU\/μg\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\u003e20mM Tris-HCl pH-8, 1mM DTT, 100mM NaCl and 10% glycerol\u003cbr\u003e\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\u003eStable for 12 months upon stored at -80℃ from the date of receipt. And avoid repeated freeze-thaws cycles.\u003cbr\u003e\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\u003eMAT1A (methionine adenosyltransferase 1A) is mainly expressed in the liver, and the protein it encodes exists as dimers or tetramers, with each subunit containing approximately 395 amino acids and possessing a typical Rossmann-fold nucleotide-binding domain and a catalytic domain. Functionally, MAT1A catalyzes the synthesis of S-adenosylmethionine (SAM) from methionine and ATP, serving as a core enzyme in hepatic one-carbon metabolism and methylation reactions. Clinically, MAT1A is often silenced in hepatocellular carcinoma and cholangiocarcinoma due to promoter hypermethylation, leading to SAM depletion, DNA hypomethylation, and genomic instability that promote tumor progression; the expression ratio of MAT1A to MAT2A can serve as a prognostic marker for liver cancer, and restoring MAT1A expression or supplementing SAM holds therapeutic potential. In addition, MAT1A mutations can cause Mudd disease (hypermethioninemia), while alcohol promotes its degradation through phosphorylation and SUMOylation, contributing to the pathogenesis of alcoholic liver disease.\u003cbr\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\u003eThe MAT1A adenosyltransferase activity is detected in a colorimetric assay wherein the monophosphate generated from the enzyme reaction forms a colored complex with the Malachite Green Phosphate Detection Kit. The amount of complex can be then measured by a microplate reader.\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\u003eMAT1A Dilution Buffer: 20mM Tris-HCl, pH8.0\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003eMAT1A Assay Buffer (5X): 100mM Tris-HCl, pH8.0, 20mM MgCl\u003csub\u003e2\u003c\/sub\u003e, 150mM KCl, 1mM DTT\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\u003c\/span\u003eMAT1A Assay Buffer (1X): 20mM Tris-HCl, pH8.0, 4mM MgCl\u003csub\u003e2\u003c\/sub\u003e, 30mM KCl, 200uM DTT\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4.\u003c\/span\u003eMAT1A His Tag Protein, Human (Catalog # UA085073)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5.\u003c\/span\u003eMalachite Green Phosphate Detection Kit (R\u0026amp;D, Catalog # DY996)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6.\u003c\/span\u003eSubstrate Assay Solution: ATP + L-methionine (Sigma, Catalog # M9625)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e7.\u003c\/span\u003e96-well half-area black plate with clear bottom (GENEVER, Catalog # GM04-96M)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e8.\u003c\/span\u003ePlate Reader (PerkinElmer)\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\u003ePrepare a Phosphate Standard curve by adding 10 µL of the 1 M Phosphate Standard to 990 µL of MAT1A Assay Buffer (1X) for 10 mM stock. Continue by adding 10 µL of the 10 mM phosphate stock to 990 µL of MAT1A Assay Buffer (1X) for 100 µM stock. (This is the first dilution to use as a standard).\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003ePerform six additional one-half dilutions of the 100 µM Phosphate stock.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\u003c\/span\u003eLoad 25 µL of each standard dilution to empty wells of a clear plate. Include a curve blank containing 25 µL MAT1A Assay Buffer (1X).  The standard curve has a range of 0.039-2.5 nmol per well.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4.\u003c\/span\u003eThaw the Active MAT1A on ice. Prepare serial dilutions of MAT1A (40 µg\/mL, 20 µg\/mL, 10 µg\/mL) using MAT1A Dilution Buffer.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5.\u003c\/span\u003eSubstrate Assay Solution: ATP solution, 10 mM; L-methionine, 150 mM.\u003c\/p\u003e\n\u003cp\u003ePrepare ATP and L-methionine diluted in distilled water to a working concentration of 2.5 mM ATP \/ 5 mM L-methionine, resulting in a final concentration of 1 mM ATP \/ 2 mM L-methionine.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6.\u003c\/span\u003eAdd the following reaction components to bring the total volume to 25 µL:\u003c\/p\u003e\n\u003cp\u003eComponent 1: 10 µL of diluted Active MAT1A (Catalog # UA085073).\u003c\/p\u003e\n\u003cp\u003eComponent 2: 10 µL of Substrate Assay Solution.\u003c\/p\u003e\n\u003cp\u003eComponent 3. 5 µL 5X MAT1A Assay Buffer\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e7.\u003c\/span\u003eSet up the blank control as outlined in step 6, excluding the addition of the Active Enzyme. Replace the MAT1A with an equal volume of MAT1A Dilution Buffer.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e8.\u003c\/span\u003eCover the plate with a plate seal and incubate the mixture in a 37℃ incubator for 60 minutes.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e9.\u003c\/span\u003eAfter incubation, add 30 µL of Malachite Green Reagent A to all wells used. Mix briefly.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e10.\u003c\/span\u003eAdd 100 μL of deionized water to all wells. Mix briefly.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e11.\u003c\/span\u003eAdd 30 µL of Malachite Green Reagent B to all wells. Mix and incubate at room temperature for 20 minutes.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e12.\u003c\/span\u003eRead the plate at 620 nm in endpoint mode.\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse:collapse;width:auto\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003eSpecific Activity (nmol\/min\/mg) =\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border-bottom:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003eCorresponding phosphate released (nmol) - Blank\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\u003ctd style=\"text-align:center;vertical-align:middle;border-top:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003eIncubation time (min) × amount of enzyme (mg)\u003c\/p\u003e\u003c\/td\u003e\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\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\u003eThe adenosyltransferase activity of MAT1A is detected by a colorimetric method. The monophosphate generated during the enzymatic reaction can form a colored complex with the Malachite Green Phosphate Detection Kit reagent, and the amount of this complex can be measured using a microplate reader.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003eExperimental materials\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003eMAT1A Dilution Buffer: 20 mM Tris-HCl, pH 8.0\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003eMAT1A Assay Buffer (5X): 100 mM Tris-HCl, pH 8.0, 20 mM MgCl₂\u003csub\u003e2\u003c\/sub\u003e, 150 mM KCl, 1 mM DTT\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\u003c\/span\u003eMAT1A Assay Buffer (1X): 20 mM Tris-HCl, pH 8.0, 4 mM MgCl₂\u003csub\u003e2\u003c\/sub\u003e, 30mM KCl, 200uM DTT\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4.\u003c\/span\u003eMAT1A His Tag Protein, Human (Catalog # UA085073)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5.\u003c\/span\u003eMalachite Green Phosphate Detection Kit (R\u0026amp;D, Catalog # DY996)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6.\u003c\/span\u003eSubstrate: ATP + L-methionine (Sigma, Catalog # M9625)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e7.\u003c\/span\u003e96-well half-area black plate with clear bottom (GENEVER, Catalog # GM04-96M)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e8.\u003c\/span\u003ePlate reader (PerkinElmer)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003eExperimental procedure\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003ePrepare the phosphate standard curve: Add 10 µL of 1 M phosphate standard solution to 990 µL of MAT1A assay buffer (1X) to prepare a 10 mM stock solution. Then, add 10 µL of the 10 mM phosphate stock solution to 990 µL of MAT1A assay buffer (1X) to prepare a 100 µM stock solution. (This serves as the first-level dilution used as the standard.)\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2.\u003c\/span\u003ePerform six serial half-fold dilutions of the 100 µM phosphate stock solution.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3.\u003c\/span\u003eTake 25 µL of each concentration of the standard dilutions and add them to the wells of a clear microplate. Also set up a blank well containing 25 µL of MAT1A assay buffer (1X). The concentration range of the standard curve is 0.039–2.5 nmol per well.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4.\u003c\/span\u003eThaw the active MAT1A on ice. Prepare a series of dilutions of MAT1A (40 µg\/mL, 20 µg\/mL, 10 µg\/mL) using the MAT1A dilution buffer.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5.\u003c\/span\u003eSubstrate detection solution: ATP solution, 10 mM; L-methionine, 150 mM.\u003c\/p\u003e\n\u003cp\u003eDilute ATP and L-methionine with distilled water to achieve a working concentration of 2.5 mM ATP \/ 5 mM L-methionine, resulting in final concentrations of 1 mM ATP \/ 2 mM L-methionine.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6.\u003c\/span\u003eAdd the following reaction components to bring the total volume to 25 µl:\u003c\/p\u003e\n\u003cp\u003eComponent 1: 10 µl of diluted active MAT1A (Cat. No. UA085073)\u003c\/p\u003e\n\u003cp\u003eComponent 2: 10 µl of substrate detection solution\u003c\/p\u003e\n\u003cp\u003eComponent 3. 5 µl of 5X MAT1A assay buffer\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e7.\u003c\/span\u003eSet up the blank control as described in step 6, but do not add MAT1A. Replace MAT1A with an equal volume of MAT1A dilution buffer.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e8.\u003c\/span\u003eCover the microplate with a plate lid and incubate the mixture in a 37°C incubator for 60 minutes.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e9.\u003c\/span\u003eAfter incubation, add 30 µL of Malachite Green Reagent A to all wells used, and mix gently.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e10.\u003c\/span\u003eAdd 100 µL of deionized water to all wells, and mix gently.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e11.\u003c\/span\u003eAdd 30 µL of Malachite Green Reagent B to all wells. After mixing, incubate at room temperature for 20 minutes.\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e12.\u003c\/span\u003eRead the microplate in endpoint mode at a wavelength of 620 nm.\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse:collapse;width:auto\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003eSpecific Activity (nmol\/min\/mg) =\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border-bottom:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003eCorresponding phosphate released (nmol) - Blank\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\u003ctd style=\"text-align:center;vertical-align:middle;border-top:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003eIncubation time (min) × amount of enzyme (mg)\u003c\/p\u003e\u003c\/td\u003e\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"10μg","offer_id":67221644247115,"sku":"UA085073-10μg","price":270.0,"currency_code":"USD","in_stock":true},{"title":"50μg","offer_id":67221644279883,"sku":"UA085073-50μg","price":540.0,"currency_code":"USD","in_stock":true},{"title":"100μg","offer_id":67221644312651,"sku":"UA085073-100μg","price":860.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/0e2e3ea4-f6ef-435d-be5b-0ddf1e42d4bb_6e0a0c84-ccbe-42a5-9d5c-5776b5c0b09c.jpg?v=1791774038","url":"https:\/\/www.antbioinc.com\/products\/mat1a-his-tag-protein-human-ua085073","provider":"AntBio","version":"1.0","type":"link"}