{"product_id":"ua-glo-nadph-detection-system-ua079042","title":"UA-Glo® NAD(P)H Detection System","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\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\u003eDry ice shipping. Upon receipt, store the kit protected from light at -80°C, valid for 12 months. After initial use, aliquot the reagents and store protected from light. Store the reductase substrate at -80°C, and the luciferin detection solution and reductase at -20°C or below. For long-term storage (\u0026gt;3 months), keep at -80°C and avoid repeated freeze-thaw cycles.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan\u003eNADH and NADPH (collectively referred to as NAD(P)H) are reactants or products in many enzymatic reactions, and the quantitative measurement of NAD(P)H can directly or indirectly determine the activity of numerous enzymes. The UA-Glo® NAD(P)H Detection Kit utilizes a coupled reaction of reductase and luciferase, where reductase equivalently consumes NADH or NADPH as substrates. Through NAD(P)H, the reductase substrate is converted into luciferin, which is then acted upon by luciferase to generate luminescence. The luminescence intensity is proportional to the amount of NAD(P)H consumed or produced in the enzymatic reaction in vitro, and this coupled reaction does not detect or is affected by the oxidized form of NAD(P).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eThe UA-Glo® NAD(P)H Detection Reagent is a homogeneous reagent, requiring only a single addition for detection, thereby reducing experimental errors caused by multiple operations. This reagent features high signal-to-noise ratio, excellent reproducibility, and stability. Its stable glow signal (half-life \u0026gt;2 hours) makes it particularly suitable for high-throughput screening of enzyme inhibitors.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eUA-Glo®\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e  \u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003eNAD(P)H\u003c\/span\u003e\u003cspan\u003e Detection Kit components and specifications are as follows. Based on an enzymatic reaction volume of \u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL for a 96-well plate and \u003c\/span\u003e\u003cspan\u003e25\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL for a 384-well plate, \u003c\/span\u003e\u003cspan\u003e10mL\u003c\/span\u003e\u003cspan\u003e allows for \u003c\/span\u003e\u003cspan\u003e200\u003c\/span\u003e\u003cspan\u003e reactions (96-well plate) or \u003c\/span\u003e\u003cspan\u003e400\u003c\/span\u003e\u003cspan\u003e reactions (384-well plate), while \u003c\/span\u003e\u003cspan\u003e50mL\u003c\/span\u003e\u003cspan\u003e allows for \u003c\/span\u003e\u003cspan\u003e1,000\u003c\/span\u003e\u003cspan\u003e reactions (96-well plate) or \u003c\/span\u003e\u003cspan\u003e2,000\u003c\/span\u003e\u003cspan\u003e reactions (384-well plate).\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eSpecification\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eLuciferin Detection Reagent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eReductase\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eReductase Substrate\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e10mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e10 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e200 \u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e50 \u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"\"\u003e\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003eReagent Preparation\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan\u003eUA-Glo®\u003c\/span\u003e\u003cspan\u003e  \u003c\/span\u003e\u003cspan\u003eNAD(P)H \u003c\/span\u003e\u003cspan\u003edetection reagent should be freshly prepared as needed. It is not recommended to store pre-prepared detection reagent. During detection, an equal volume of detection reagent as the enzymatic reaction volume should be added. Calculate the required volume of detection reagent accordingly.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan\u003eTake out the three components from the \u003c\/span\u003e\u003cspan\u003eUA-Glo®\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e   \u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003eNAD(P)H\u003c\/span\u003e\u003cspan\u003e detection kit.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003eAfter thawing the luciferin detection solution at room temperature, it can be kept at room temperature\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e(22-25°C) \u003c\/span\u003e\u003cspan\u003euntil use\u003c\/span\u003e\u003cspan\u003e(\u003c\/span\u003e\u003cspan\u003enot exceeding\u003c\/span\u003e\u003cspan\u003e24hr)\u003c\/span\u003e\u003cspan\u003e, or stored at\u003c\/span\u003e\u003cspan\u003e4°C\u003c\/span\u003e\u003cspan\u003efor up to one week. For long-term storage, aliquot and store at\u003c\/span\u003e\u003cspan\u003e-20°C\u003c\/span\u003e\u003cspan\u003eor lower temperatures, avoiding repeated freeze-thaw cycles.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003eThe reductase concentration is\u003c\/span\u003e\u003cspan\u003e50x\u003c\/span\u003e\u003cspan\u003e. After thawing, add the required volume (see step\u003c\/span\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003e) to the luciferin detection solution. The remaining reductase should be aliquoted and stored at\u003c\/span\u003e\u003cspan\u003e-20°C\u003c\/span\u003e\u003cspan\u003eor lower temperatures, avoiding repeated freeze-thaw cycles.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan\u003eThe reductase substrate concentration is\u003c\/span\u003e\u003cspan\u003e200x\u003c\/span\u003e\u003cspan\u003e. After thawing, add the required volume (see step\u003c\/span\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003e) to the luciferin detection solution.\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eThe remaining reductase substrate should be aliquoted and stored at\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e-80°C\u003c\/span\u003e\u003cspan\u003e, avoiding repeated freeze-thaw cycles.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan\u003eNAD(P)H\u003c\/span\u003e\u003cspan\u003edetection reagent preparation: The reductase concentration is\u003c\/span\u003e\u003cspan\u003e50x\u003c\/span\u003e\u003cspan\u003e, and the reductase substrate concentration is\u003c\/span\u003e\u003cspan\u003e200x\u003c\/span\u003e\u003cspan\u003e. Mix the two components into the luciferin detection solution at this ratio to prepare the\u003c\/span\u003e\u003cspan\u003eNAD(P)H\u003c\/span\u003e\u003cspan\u003edetection reagent. For example, add\u003c\/span\u003e\u003cspan\u003e20\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eof reductase and\u003c\/span\u003e\u003cspan\u003e5\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eof reductase substrate to\u003c\/span\u003e\u003cspan\u003e1mL\u003c\/span\u003e\u003cspan\u003eof luciferin detection solution. Invert the reagent to mix. It is recommended to use the prepared\u003c\/span\u003e\u003cspan\u003eNAD(P)H\u003c\/span\u003e\u003cspan\u003edetection reagent immediately.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNAD(P)H \u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eDetection\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan\u003ePerform enzymatic reactions that consume or generate\u003c\/span\u003e\u003cspan\u003eNAD(P)H)\u003c\/span\u003e\u003cspan\u003eaccording to experimental requirements:\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003ea)\u003c\/span\u003e\u003cspan\u003eIt is recommended to use white opaque assay plates for enzymatic reactions. The reaction volume for a 96-well plate is\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003e, for a 384-well plate is\u003c\/span\u003e\u003cspan\u003e25\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003e, or other appropriate enzymatic reaction volumes. During detection, add an equal volume of\u003c\/span\u003e\u003cspan\u003eUA-Glo®\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e  \u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003eNAD(P)H\u003c\/span\u003e\u003cspan\u003edetection reagent as the enzymatic reaction volume.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003eb)\u003c\/span\u003e\u003cspan\u003eAvoid reducing agents such as\u003c\/span\u003e\u003cspan\u003eDTT\u003c\/span\u003e\u003cspan\u003ein enzymatic reactions. Reducing agents may react with the reductase substrate and increase detection background.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003eBefore\u003c\/span\u003e\u003cspan\u003eNAD(P)H \u003c\/span\u003e\u003cspan\u003edetection, equilibrate the assay plate to room temperature. The luciferase reaction is sensitive to temperature changes. Both the detection reagent and the assay plate need to be equilibrated to room temperature, and the temperature should remain constant during testing\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e(±1\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003e).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003eAdd an equal volume of\u003c\/span\u003e\u003cspan\u003eUA-Glo®\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e  \u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003eNAD(P)H\u003c\/span\u003e\u003cspan\u003edetection reagent as the enzymatic reaction volume to each well of the assay plate.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan\u003eShake the plate at medium speed for\u003c\/span\u003e\u003cspan\u003e30s\u003c\/span\u003e\u003cspan\u003e, then incubate in the dark for\u003c\/span\u003e\u003cspan\u003e40-60min\u003c\/span\u003e\u003cspan\u003eto stabilize the luminescent signal.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan\u003eRead the luminescent signal on a luminometer or multifunctional plate reader. The plate can be read within\u003c\/span\u003e\u003cspan\u003e2-3hr\u003c\/span\u003e\u003cspan\u003e. The luminescent signal typically peaks at\u003c\/span\u003e\u003cspan\u003e60min\u003c\/span\u003e\u003cspan\u003e, with a signal half-life of\u003c\/span\u003e\u003cspan\u003e\u0026gt;2hr\u003c\/span\u003e\u003cspan\u003e.\u003c\/span\u003e\u003cspan\u003eNAD(P)H\u003c\/span\u003e\u003cspan\u003edetection reagent stops most enzymatic reactions upon addition. However, some enzymes may continue to react after the addition of the detection reagent. If a stable signal is required, add a specific inhibitor for such enzymes to the\u003c\/span\u003e\u003cspan\u003eNAD(P)H\u003c\/span\u003e\u003cspan\u003edetection reagent before adding it to the enzymatic reaction for detection.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e1) Different batches are not recommended for mixed use  2) Without strict validation, it is not recommended to change the dosage of detection reagents  3) For research use only\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"10ml","offer_id":43093874769995,"sku":"UA079042-10ml","price":220.0,"currency_code":"USD","in_stock":true},{"title":"50ml","offer_id":43093874802763,"sku":"UA079042-50ml","price":930.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_19ad7041-710f-4692-856a-5de76261c556.png?v=1788199372","url":"https:\/\/www.antbioinc.com\/products\/ua-glo-nadph-detection-system-ua079042","provider":"AntBio","version":"1.0","type":"link"}