bSLSBio LifeSpan BioSciences, Inc. Lactate Dehydrogenase (LDH) Assay Kit (Colorimetric) LS-K306-100 (100 Tests) • Store at -20°C Introduction LACTATE DEHYDROGENASE (LDH) is an oxidoreductase which catalyzes the interconversion of lactate and pyruvate. When disease or injury affects tissues containing LDH, the cells release LDH into the bloodstream, where it is identified in higher than normal levels. Therefore, LDH is most often measured to evaluate the presence of tissue or cell damage. The nonradioactive colorimetric LDH assay is based on the reduction of the tetrazolium salt MTT in a NADH-coupled enzymatic reaction to a reduced form of MTT which exhibits an absorption maximum at 565 nm. The intensity of the purple color formed is directly proportional to the enzyme activity. Key Features • High sensitivity and wide linear range. Use 3 pL serum or plasma sample. The detection limit is 2 U/L, linear up to 200 U/L. • Homogeneous and simple procedure. Simple "mix-and-measure" procedure allows reliable quantitation of LDH activity within 30 minutes. • Robust and amenable to HTS. All reagents are compatible with high-throughput liquid handling instruments. Applications • Direct Assays: LDH activity in serum, plasma and other sources. • Characterization and Quality Control for LDH production. • Drug Discovery: screen and evaluation of LDH modulators. Materials Not Supplied Pipetting devices and accessories (e.g. multi-channel pipettor), clear flat-bottom 96-well plates (e.g. VWR cat# 82050-760) or cuvettes and plate reader or spectrophotometer capable of measuring OD 565 nm. Storage The kit is shipped at ambient temperature. Store all components at -20°C upon receiving. Shelf life: 6 months after receipt. FOR RESEARCH USE ONLY! Not for use in humans. LifeSpan BioSciences, Inc. • 2401 Fourth Avenue, Suite 900, Seattle, WA 98121 www.LSBio.com • (206) 464-1554 • [email protected]

bSLSBio LifeSpan BioSciences, Inc. Lactate Dehydrogenase (LDH) Assay Kit (Colorimetric) LS-K306-100 (100 Tests) • Store at -20°C Assay Procedure This assay is based on a kinetic reaction. To ensure identical incubation time, addition of Working Reagent to samples should be quick and mixing should be brief but thorough. Use of a multi-channel pipettor is recommended. Assays can be executed at room temperature or 30°C. Sample Preparation Serum and plasma are assayed directly. Tissue: prior to dissection, rinse tissue in phosphate buffered saline (pH 7.4) to remove blood. Homogenize tissue in 5...

bSLSBio LifeSpan BioSciences, Inc. Lactate Dehydrogenase (LDH) Assay Kit (Colorimetric) LS-K306-100 (100 Tests) • Store at -20°C Sample Data Samples were assayed using the 96-well plate protocol. The LDH activity (IU/L) was 41 for a human serum, 220 for rat serum and 88 for fetal bovine serum, respectively. Kinetics of LDH reaction in 96-well plate assay with increasing serum concentration FOR RESEARCH USE ONLY! Not for use in humans. LifeSpan BioSciences, Inc. • 2401 Fourth Avenue, Suite 900, Seattle, WA 98121 www.LSBio.com • (206) 464-1554 • [email protected]