{"product_id":"ldh-colorimetry-cytotoxicity-assay-ua079060","title":"LDH Colorimetry Cytotoxicity Assay","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\u003e\u003cspan\u003eLDH assay reagents and the LDH positive control should be shipped on dry ice, while the stop solution (1N HCl) and cell lysis buffer (10x) should be shipped with ice packs. Upon receipt, store the LDH assay reagents and the LDH positive control protected from light at -80°C, and keep the stop solution (1N HCl) and cell lysis buffer (10x) at 4°C. Refer to the reagent vial labels for the expiration dates. After first use, it is recommended to aliquot the LDH assay reagents as needed and store them protected from light at -20°C or below; for long-term storage (\u0026gt;2 months), store at -80°C. The LDH assay reagents should not be subjected to more than three freeze-thaw cycles after initial use. For the LDH positive control, after first use, we recommend long-term storage by aliquoting as needed and keeping at -20°C or below; for short-term storage (up to 2 weeks), store at 4°C.\u003c\/span\u003e\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\u003cp\u003e\u003cspan\u003eThe LDH Colorimetric Cytotoxicity Assay Kit determines the toxicity of external stimuli or compounds to cells by measuring the lactate dehydrogenase (LDH) released by cells. LDH is widely present in the cytoplasm and is released into the culture medium when cell membranes are damaged or ruptured. With a half-life of approximately 9 hours in the culture medium, extracellular LDH is often used as an indicator of cell health to reflect the toxic effects of external stimuli or compounds. The LDH colorimetric cytotoxicity assay detects LDH activity through an enzymatic reaction. Released LDH converts NAD to NADH, and a reductase then uses NADH to convert iodonitrotetrazolium violet (INT) into a red formazan product. The absorbance of the formazan product can be measured at 490 nm (or 492 nm) for quantification, with its amount being directly proportional to the LDH activity in the sample. The LDH colorimetric cytotoxicity assay kit features a high signal-to-noise ratio, good reproducibility, and excellent stability. Its ready-to-use homogeneous formulation requires only a single addition step, reducing experimental errors that may arise from multiple additions. Additionally, this reagent does not require cell lysis, allowing continuous testing on the same cell sample and further simplifying experimental design. The LDH colorimetric cytotoxicity assay kit can be used in combination with other assays for assessing cell health on the same experimental sample, such as the UA-Glo® CTG Cell Viability Assay Kit (Cat# UA070103), Caspase\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eA cell apoptosis assay kit (Cat# UA079057) and the UA-Glo® fluorescent cell viability assay kit (Cat# UA079015) were used for multiplexed detection to better elucidate the mechanisms of cytotoxicity.\u003c\/span\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan\u003eThe components and specifications of the colorimetric cytotoxicity assay kit are as follows. Detectable\u003c\/span\u003e\u003cspan\u003e96\/384-\u003c\/span\u003e\u003cspan\u003ewell plate reaction numbers are calculated based on adding\u003c\/span\u003e\u003cspan\u003e50\/12.5\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμ\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eL\u003c\/span\u003e\u003cspan\u003esample and\u003c\/span\u003e\u003cspan\u003e50\/12.5\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμ\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eL\u003c\/span\u003e\u003cspan\u003edetection reagent per well.\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\"\u003eSpecifications\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\"\u003eComponents and specifications\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\"\u003e96-\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003ewell plate reaction number\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\"\u003e384-\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003ewell plate reaction number\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\"\u003e200T\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\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003eLDH\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003edetection reagent:\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e12 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003estop solution\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e(\u003c\/span\u003e\u003cspan\u003e1N HCl)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e:\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e12 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003ecell lysis buffer\u003c\/span\u003e\u003cspan\u003e(10×)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e:\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e1 ml\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003eLDH\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003epositive control\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e(500x): 50\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμ\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eL\u003c\/span\u003e\u003c\/p\u003e\n\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\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\u003e800\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\"\u003e1000T\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\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan\u003edetection reagent:\u003c\/span\u003e\u003cspan\u003e12x5 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003estop solution\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e(\u003c\/span\u003e\u003cspan\u003e1N HCl)\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003e60 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003ecell lysis buffer\u003c\/span\u003e\u003cspan\u003e(10×)\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003e5 ml\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003epositive control\u003c\/span\u003e\u003cspan\u003e(500x): 50 μL\u003c\/span\u003e\u003c\/p\u003e\n\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\u003e1,000\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\u003e4,000\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"font-weight:bold\"\u003eExperimental Control\u003c\/span\u003e\u003cspan\u003e: Under most experimental conditions, it is not recommended to directly compare the absolute readings of sample measurements. It is suggested to set up the same reference controls on each experimental plate; for example, in a tumor cell cytotoxicity assay, use the same toxic reference compound, solvent vehicle control, and medium blank control. Normalize the sample measurement values using the readings from the toxic reference compound and solvent vehicle control before data processing and comparison.\u003c\/span\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan\u003eCytotoxicity assays using the colorimetric method can calculate cytotoxicity according to the following formula:\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e(%)\u003c\/span\u003e\u003cspan\u003e: Cytotoxicity\u003c\/span\u003e\u003cspan\u003e(%) = 100x (\u003c\/span\u003e\u003cspan\u003eSample measurement value\u003c\/span\u003e\u003cspan\u003e-\u003c\/span\u003e\u003cspan\u003eSolvent vehicle control measurement value\u003c\/span\u003e\u003cspan\u003e)\/(\u003c\/span\u003e\u003cspan\u003eMaximum\u003c\/span\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan\u003erelease control measurement value\u003c\/span\u003e\u003cspan\u003e-\u003c\/span\u003e\u003cspan\u003eSolvent vehicle control measurement value\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003cspan\u003e. The maximum\u003c\/span\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan\u003erelease control can be either the toxic reference compound control or, prior to the assay,\u003c\/span\u003e\u003cspan\u003e45min\u003c\/span\u003e\u003cspan\u003ebefore adding the solvent vehicle control, add lysis buffer\u003c\/span\u003e\u003cspan\u003e(10×)\u003c\/span\u003e\u003cspan\u003eto achieve a final concentration\u003c\/span\u003e\u003cspan\u003eof 1x\u003c\/span\u003e\u003cspan\u003e, for example,\u003c\/span\u003e\u003cspan\u003e100\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eof culture medium with\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eof lysis buffer\u003c\/span\u003e\u003cspan\u003e(10×)\u003c\/span\u003e\u003cspan\u003efollowed by\u003c\/span\u003e\u003cspan\u003e45min (\u003c\/span\u003e\u003cspan\u003ei.e., simultaneously with the samples\u003c\/span\u003e\u003cspan\u003e)\u003c\/span\u003e\u003cspan\u003eCollected supernatant. It is recommended to set up a cell-free complete medium well as a background control for detection; this background reading can be used to correct the experimental readings. The kit includes one tube\u003c\/span\u003e\u003cspan\u003eLDH (500x)\u003c\/span\u003e\u003cspan\u003epositive control, which can be diluted with\u003c\/span\u003e\u003cspan\u003ePBS+1% BSA\u003c\/span\u003e\u003cspan\u003eto make a\u003c\/span\u003e\u003cspan\u003e500x\u003c\/span\u003e\u003cspan\u003edilution as the positive control for the experiment.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003eThe following example describes a compound cytotoxicity assay using a\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e96\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003ewell cell plate.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003e3.1\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eCell treatment\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan\u003eIn a\u003c\/span\u003e\u003cspan\u003e96-well cell culture plate, seed the experimental cells at an appropriate density, with\u003c\/span\u003e\u003cspan\u003e100-200\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eof culture medium per well. Serum, such as\u003c\/span\u003e\u003cspan\u003eFBS\u003c\/span\u003e\u003cspan\u003emay contain\u003c\/span\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan\u003e. If this background is high enough to affect the experimental results, it is recommended to use\u003c\/span\u003e\u003cspan\u003e56\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003e0.5 hours\u003c\/span\u003e\u003cspan\u003eto inactivate the\u003c\/span\u003e\u003cspan\u003eFBS\u003c\/span\u003e\u003cspan\u003ewhen preparing the complete culture medium for seeding the cells.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003ePerform compound treatment on the cells according to experimental requirements. Each experimental plate should include\u003c\/span\u003e\u003cspan\u003ea toxic reference compound control, a solvent vehicle control, and a\u003c\/span\u003e\u003cspan\u003ecell-free complete medium control.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003eAccording to experimental requirements, samples are taken at a certain time after cell treatment for\u003c\/span\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan\u003edetection.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003e \u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003e3.2 LDH\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\u003eTake\u003c\/span\u003e\u003cspan\u003e50 μL\u003c\/span\u003e\u003cspan\u003eof the above-mentioned culture medium sample and add it to a completely transparent\u003c\/span\u003e\u003cspan\u003e96-\u003c\/span\u003e\u003cspan\u003ewell plate. The maximum detectable value should preferably not exceed\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003ewhen reading the plate at\u003c\/span\u003e\u003cspan\u003e15 min\u003c\/span\u003e\u003cspan\u003e. If it exceeds\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003e, it is recommended to appropriately dilute the sample before detection.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003eCalculate\u003c\/span\u003e\u003cspan\u003ethe amount of LDH\u003c\/span\u003e\u003cspan\u003edetection reagent needed:\u003c\/span\u003e\u003cspan\u003eFor 96-\u003c\/span\u003e\u003cspan\u003ewell plate detection, calculate based on adding\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eµL\u003c\/span\u003e\u003cspan\u003eof sample with\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eµL of LDH\u003c\/span\u003e\u003cspan\u003edetection reagent.\u003c\/span\u003e\u003cspan\u003eFor 384-\u003c\/span\u003e\u003cspan\u003ewell plate detection, calculate based on adding\u003c\/span\u003e\u003cspan\u003e12.5\u003c\/span\u003e\u003cspan\u003eµL\u003c\/span\u003e\u003cspan\u003eof sample with\u003c\/span\u003e\u003cspan\u003e12.5\u003c\/span\u003e\u003cspan\u003eµL of LDH\u003c\/span\u003e\u003cspan\u003edetection reagent.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003eRemove\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e \u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003ethe LDH\u003c\/span\u003e\u003cspan\u003edetection reagent from storage, thaw it, and equilibrate it to room temperature\u003c\/span\u003e\u003cspan\u003e(22\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003e-25\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003e).\u003c\/span\u003e\u003cspan\u003eInvert thoroughly to mix well. After taking the required amount of reagent according to experimental needs, use it immediately; store any remaining reagent by freezing at\u003c\/span\u003e\u003cspan\u003e-20\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003eor lower.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan\u003eAdd to each well\u003c\/span\u003e\u003cspan\u003e50 µL\u003c\/span\u003e\u003cspan\u003eAdd sample\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eµL LDH\u003c\/span\u003e\u003cspan\u003edetection reagent. When handling multiple samples, use a multichannel pipette to add the detection reagent to ensure consistent incubation time.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan\u003eShake the plate at medium speed\u003c\/span\u003e\u003cspan\u003efor 15 sec\u003c\/span\u003e\u003cspan\u003eto mix thoroughly, then incubate in the dark at room temperature.\u003c\/span\u003e\u003cspan\u003eAfter\u003c\/span\u003e\u003cspan\u003e15 min\u003c\/span\u003e\u003cspan\u003eincubation,\u003c\/span\u003e\u003cspan\u003eread the absorbance at 490 nm. For low-concentration\u003c\/span\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan\u003eassays, the incubation time can be extended to\u003c\/span\u003e\u003cspan\u003e30 min\u003c\/span\u003e\u003cspan\u003eto improve detection sensitivity.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6)\u003c\/span\u003e\u003cspan\u003eAdd\u003c\/span\u003e\u003cspan\u003eLDH\u003c\/span\u003e\u003cspan\u003edetection reagent\u003c\/span\u003e\u003cspan\u003eand incubate for\u003c\/span\u003e\u003cspan\u003e15 min\u003c\/span\u003e\u003cspan\u003e(or\u003c\/span\u003e\u003cspan\u003e30 min).\u003c\/span\u003e\u003cspan\u003eAfter incubation,\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eadd\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eµL\u003c\/span\u003e\u003cspan\u003eof stop solution\u003c\/span\u003e\u003cspan\u003e(1N HCl),\u003c\/span\u003e\u003cspan\u003eshake the plate at medium speed\u003c\/span\u003e\u003cspan\u003efor 15 sec\u003c\/span\u003e\u003cspan\u003eto mix well.\u003c\/span\u003e\u003cspan\u003eRead the absorbance at\u003c\/span\u003e\u003cspan\u003e490 nm\u003c\/span\u003e\u003cspan\u003ewithin 1 hour; the signal remains essentially stable. The stop solution is\u003c\/span\u003e\u003cspan\u003e1N HCl\u003c\/span\u003e\u003cspan\u003eand should be handled with care and proper protective measures.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"margin-bottom: 0px;\"\u003eIt is not recommended to mix different batches.\u003c\/p\u003e\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"200T","offer_id":67255154540619,"sku":"UA079060-200T","price":85.0,"currency_code":"USD","in_stock":true},{"title":"1000T","offer_id":67255154573387,"sku":"UA079060-1000T","price":280.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_708039b9-029e-4e07-8e7d-f3174a995d13.png?v=1792177205","url":"https:\/\/www.antbioinc.com\/products\/ldh-colorimetry-cytotoxicity-assay-ua079060","provider":"AntBio","version":"1.0","type":"link"}