{"product_id":"nnmt-his-tag-protein-rhesus-macaque-ua085036","title":"NNMT His Tag Protein, Rhesus macaque","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%;\"\u003eRhesus macaque\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%;\"\u003eNicotinamide N-methyltransferase\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%;\"\u003eF7ERX8\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-Leu264 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;\"\u003e25-35kDa (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,＞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, 200mM NaCl, 20% Glycerol, 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.Swaminathan S, Birudukota S, Thakur MK, Parveen R, Kandan S, Juluri S, Shaik S, Anand NN, Burri RR, Kristam R, Hallur MS, Rajagopal S, Schreuder H, Langer T, Rudolph C, Ruf S, Dhakshinamoorthy S, Gosu R, Kannt A. Crystal structures of monkey and mouse nicotinamide N-methyltransferase (NNMT) bound with end product, 1-methyl nicotinamide. Biochem Biophys Res Commun. 2017 Sep 16;491(2):416-422.\u003cbr\u003e2.Parsons RB, Smith ML, Williams AC, Waring RH, Ramsden DB. Expression of nicotinamide N-methyltransferase (E.C. 2.1.1.1) in the Parkinsonian brain. J Neuropathol Exp Neurol. 2002 Feb;61(2):111-24.\u003cbr\u003e3.Kocinaj A, Chaudhury T, Uddin MS, Junaid RR, Ramsden DB, Hondhamuni G, Klamt F, Parsons L, Parsons RB. High Expression of Nicotinamide N-Methyltransferase in Patients with Sporadic Alzheimer's Disease. Mol Neurobiol. 2021 Apr;58(4):1769-1781.\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\u003eNNMT (Nicotinamide N-methyltransferase) is a cytosolic methyltransferase whose crystal structure has been resolved in rhesus macaques and mice at a resolution of 1.88 Å, belonging to the methyltransferase superfamily. This enzyme utilizes S-adenosylmethionine as a methyl donor to catalyze the methylation of nicotinamide into 1-methylnicotinamide, thereby regulating NAD⁺ metabolic homeostasis; it is highly expressed in the liver, adipose tissue, and brain, and participates in energy expenditure, lipid metabolism, and insulin sensitivity modulation through the NAD⁺\/Sirt1 pathway. Clinical studies have demonstrated that upregulated NNMT expression is closely associated with obesity, type 2 diabetes, and metabolic syndrome, while its levels are also significantly elevated in specific brain regions of patients with Parkinson's disease and Alzheimer's disease, suggesting that NNMT serves not only as a biomarker for metabolic disorders but also as a promising therapeutic target for drug intervention.\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\u003eThe MTase-Glo™ Methyltransferase Assay can measure the activity of NNMT based on the reaction product S-adenosyl homocysteine (SAH). After the methyltransferase reaction is complete, just add the MTase-Glo™ Reagent and MTase-Glo™ Detection Solution to convert SAH to ATP and measure the light from the luciferase 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\u003eAssay Buffer: 20mM Tris-HCl, pH 8.0, 50 mM NaCl, 1 mM EDTA, 3 mM MgCl\u003csub\u003e2\u003c\/sub\u003e, 0.1 mg\/ml BSA. Add 1mM DTT prior to use.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e2.\u003c\/span\u003e\u003cspan\u003eNNMT His tag Protein, Rhesus macaque\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e3.\u003c\/span\u003e\u003cspan\u003eMTase-Glo\u003csup\u003eTM\u003c\/sup\u003e Methyltransferase Assay (Promega, Catalog #: V7601)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e4.\u003c\/span\u003e\u003cspan\u003eSubstrate: Nicotinamide (Sigma, Catalog #: N481907-100GM)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right:8px;\"\u003e5.\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;\"\u003e6.\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\u003e1. Thaw the active NNMT and all Methyltransferase-Glo\u003csup\u003eTM\u003c\/sup\u003e Assays kit reagents on ice.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e2. Prepare the following working solutions with Methyltransferase Reaction Buffer on ice:\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e2X final concentration of Active NNMT: 0.4 μg\/mL, 0.2 μg\/mL, 0.1 μg\/mL, 0.05 μg\/mL, 0 μg\/mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e2X Substrate Cocktail: 40 μM of SAM and 80 μM of Nicotinamide in Reaction Buffer\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e3. In a 384-well plate, add the following components to bring the initial reaction volume to 20 μL:\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003eComponent 1. 10 μL of 2X Substrate Cocktail\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003eComponent 2. 10 μL of 2X Active NNMT\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e4. Mix the reaction on an orbital shaker for 2 minutes. Seal the plate with a plate seal and incubate at 37°C for 60 minutes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e5. Dilute 10X Methyltransferase-Glo\u003csup\u003eTM\u003c\/sup\u003e Reagent with equal volume of nanopure water, and add 5 μL of the 5X Methyltransferase-Glo\u003csup\u003eTM\u003c\/sup\u003e Reagent to all reaction wells.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e6. Mix on an orbital shaker for 2 minutes and then incubate at room temperature for 30 minutes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e7. Add 25 μL of MTase-Glo\u003csup\u003eTM\u003c\/sup\u003e Detection Solution to all reaction wells. Mix for 2 minutes and then incubate at room temperature for 30 minutes\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e8. Read at luminescence, respectively in endpoint mode.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left;\"\u003e\u003cspan\u003e9. Using the SAH standard curve, determine the concentration of SAH produced (pmol) and calculate the methyltransferase specific activity as outlined below.\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\u003eSAH (pmol)-Blank\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 \u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"10μg","offer_id":43094329655371,"sku":"UA085036-10μg","price":470.0,"currency_code":"USD","in_stock":true},{"title":"50μg","offer_id":43094329688139,"sku":"UA085036-50μg","price":1400.0,"currency_code":"USD","in_stock":true},{"title":"100μg","offer_id":43094329720907,"sku":"UA085036-100μg","price":2240.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/323b3189009444b7ad3e1a55c8af4a3f.jpg?v=1788206411","url":"https:\/\/www.antbioinc.com\/products\/nnmt-his-tag-protein-rhesus-macaque-ua085036","provider":"AntBio","version":"1.0","type":"link"}