QuantiChrom™ Lactate Dehydrogenase Kit

Protocol SDS

Application

• For quantitative determination of lactate dehydrogenase LDH activity and screen/evaluation of LDH modulators.

Key Features

• High sensitivity and wide linear range. Use 3 µL serum or plasma sample. The detection limit is 2 U/L, linear up to 200 U/L.
• Homogeneous and simple procedure. A 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.

Method

• OD565nm

Samples

• Serum, plasma etc

Species

• All

Procedure

• 30 min

Size

• 100 tests

Detection Limit

• 2 U/L

Shelf Life

• 6 months

More Details

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 at higher than normal levels. Therefore, LDH is most often measured to evaluate the presence of tissue or cell damage. The non-radioactive colorimetric LDH assay is based on the reduction of the tetrazolium salt MTT in an 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.

Is there a difference in the values of samples that are run fresh (never frozen) and samples that have been frozen and thawed (single thaw)?

LDH activity in serum samples remains relatively stable if stored at -20°C, and there is no significant difference between fresh and frozen serum.

Does hemolysis affect the assay?

Hemolysis will affect the assay itself significantly and it will add a background caused by hemoglobin that may lower the sensitivity of the assay. Hemolysis will also change your sample drastically by releasing LDH from red blood cells into the serum. Thus, you will be measuring both, the regular LDH activity in serum, as well as the LDH activity caused by hemolysis. In fact, LDH can be used as a marker for hemolysis. If you want to determine in the LDH activity in serum, you must prevent hemolysis.

Is it possible to take down the substrate buffer solution in each well so that we could use more than 10 µL of our sample?

Yes, you can use (up to) 50 µL sample instead of 10 µL, plus 30 µL color reagent and 120 µL substrate buffer, if your sample has low LDH activity. This will increase the sensitivity of this assay.

Is this assay for lactate dehydrogenase activity affected by which anticoagulant is used in the blood draw?

Citrate, EDTA, and heparin, are all compatible with the QuantiChrom Lactate dehydrogenase assay kit when used as an anticoagulant for blood collection

Gabel, M et al (2020). Annexin A2 Egress during Calcium-Regulated Exocytosis in Neuroendocrine Cells. Cells, 9(9), E2059. Assay: lactate dehydrogenase in bovine chromaffin cells.

Ryan, E. L., Klopfenstein, J. J., & Kutzler, M. A. (2020). Intramammary antibiotics with complementary acupuncture decreases milk serum N-acetyl-beta-D-glucosaminidase concentrations in dairy cattle with subclinical mastitis. Reproduction in Domestic Animals, 55(12), 1747-1755. Assay: lactate dehydrogenase in bovine milk serum.

Charrin, E et al (2018). Moderate exercise training decreases inflammation in transgenic sickle cell mice. Blood Cells, Molecules and Diseases. 69:45-52. Assay: Lactate dehydrogenase in mice cell and sickle.

Haney, SL et al (2018). Preclinical investigation of a potent geranylgeranyl diphosphate synthase inhibitor. Investigational New Drugs. 36(5): 810-818. Assay: Lactate dehydrogenase in mice cells.

Berlingieri, S. (2017). Gain And Loss Of Progranulin Have Opposite Effects On Autophagy (Doctoral dissertation, The Open University). Assay: Lactate dehydrogenase in human liver cells.

Sharabi, K. et al (2017). Selective chemical inhibition of PGC-1alpha gluconeogenic activity ameliorates type 2 diabetes. Cell, 169(1), 148-160. Assay: Lactate dehydrogenase in mice liver cells.

Ghosh, Samit, et al. (2016). Nonhematopoietic Nrf2 dominantly impedes adult progression of sickle cell anemia in mice. JCI insight 1(4). Assay: Lactate dehydrogenase in mice plasma.

Higashiyama, M et al (2012).Indomethacin-induced small intestinal injury is ameliorated by cilostazol, a specific PDE-3 inhibitor. Scandanavian Journal of Gastroenterology 47(8-9):993-1002. Assay: Lactate dehydrogenase in mice intestinal cells.

Demers, M., et al. (2011). Increased efficacy of breast cancer chemotherapy in thrombocytopenic mice. Cancer Res 71(5):1540-9. Assay: Lactate dehydrogenase in mouse tumor homogenate.

Farina, F., et al. (2011). The acute toxic effects of particulate matter in mouse lung are related to size and season of collection. Toxicol Lett 202(3):209-17. Assay: Lactate dehydrogenase in mouse BALF.

Heo, Y.J., et al. (2011). The expression of the receptor for advanced glycation end-products (RAGE) in RA-FLS is induced by IL-17 via Act-1. Arthritis Res Ther 13(4):R113. Assay: Lactate dehydrogenase in human synoviocyte culture supernatant.

Lazado, C.C., et al. (2011). In vitro adherence of two candidate probiotics from Atlantic cod and their interference with the adhesion of two pathogenic bacteria. Vet Microbiol 148(2-4):252-9. Assay: Lactate dehydrogenase in atlantic cod intestinal cell supernatant.

Meshulam, T., et al. (2011). Caveolins/caveolae protect adipocytes from fatty acid-mediated lipotoxicity. J Lipid Res 52(8):1526-32. Assay: Lactate dehydrogenase in mouse cell supernatant.

Methods for treatment of oncological disorders using an epimetabolic shifter (coenzyme q10). US Patent Appl. 20110027247. Assay: Lactate Dehydrogenase in mouse cell.

Pantos, C., et al. (2011). Acute T3 treatment protects the heart against ischemia-reperfusion injury via TRalpha1 receptor. Mol Cell Biochem 353(1-2):235-41. Assay: Lactate dehydrogenase in rat heart perfudate.

Uchida, S., et al. (2011). Combination of chondroitin sulfate and polyplex micelles from Poly(ethylene glycol)-poly{N’-[N-(2-aminoethyl)-2-aminoethyl]aspartamide} block copolymer for prolonged in vivo gene transfection with reduced toxicity. J Control Release 155(2):296-302. Assay: Lactate dehydrogenase in mouse BALF.

Kulik, TB et al (2010). The relationship between oxygen and adenosine in astrocytic cultures. Glia 58(11):1335-44. Assay: Lactate dehydrogenase in rat primary astrocytes, culture medium.

Schmelzer, E., et al. (2010). Three-dimensional perfusion bioreactor culture supports differentiation of human fetal liver cells. Tissue Eng Part A 16(6):2007-16. Assay: Lactate dehydrogenase in human bioreactor medium.

Stentz, R., et al. (2010). Controlled release of protein from viable Lactococcus lactis cells. Appl Environ Microbiol 76(9):3026-31. Assay: Lactate dehydrogenase in Lactococcus lactis bacteria culture medium.

Tanaka, T., et al. (2010). In vivo evaluation of safety of nanoporous silicon carriers following single and multiple dose intravenous administrations in mice. Int J Pharm 402(1-2):190-7. Assay: Lactate dehydrogenase in mouse plasma.

van Vlerken, L.E., et al. (2010). Augmentation of therapeutic efficacy in drug-resistant tumor models using ceramide coadministration in temporal-controlled polymer-blend nanoparticle delivery systems. AAPS J 12(2):171-80. Assay: Lactate dehydrogenase in mouse plasma.

Blanco Garcia, J., et al. (2009). Physiopathological effects of the NO donor 3-morpholinosydnonimine on rat cortical synaptosomes. Neurochem Res 34(5):931-41. Assay: Lactate dehydrogenase in rat synaptosomal suspension.

Branciari, R., et al. (2009). Effect of genotype and rearing system on chicken behavior and muscle fiber characteristics. J Anim Sci 87(12):4109-17. Assay: Lactate dehydrogenase in chicken serum.

Cirrik, S, Oner, G. (2009). The effect of heavy muscle activity on renal cytoresistance in rats. Ren Fail 31(8):683-9. Assay: Lactate dehydrogenase in rat blood.

Kim, Y.M., et al. (2009). Palmatine from Coptidis rhizoma reduces ischemia-reperfusion-mediated acute myocardial injury in the rat. Food Chem Toxicol 47(8):2097-102. Assay: Lactate dehydrogenase in rat serum.

Kliment, C.R., et al. (2009). Extracellular superoxide dismutase regulates cardiac function and fibrosis. J Mol Cell Cardiol 47(5):730-42. Assay: Lactate dehydrogenase in mouse serum.

Mittal, R., et al. (2009). Brain damage in newborn rat model of meningitis by Enterobacter sakazakii: a role for outer membrane protein A. Lab Invest 89(3):263-77. Assay: Lactate dehydrogenase in rat serum, brain , intestine.

Yu, C.H., et al. (2009). Inhibitory mechanisms of Agaricus blazei Murill on the growth of prostate cancer in vitro and in vivo. J Nutr Biochem 20(10):753-64. Assay: Lactate dehydrogenase in mouse serum.

Lu, F., et al. (2008). The effect of C1 inhibitor on intestinal ischemia and reperfusion injury. Am J Physiol Gastrointest Liver Physiol 295(5):G1042-9. Assay: Lactate dehydrogenase in mouse serum.

Marques, R., et al. (2008). B lymphocyte activation by coinfection prevents immune control of friend virus infection. J Immunol 181(5):3432-40. Assay: Lactate dehydrogenase in mouse serum.

Moertel, L., et al. (2008). Comparative real-time PCR and enzyme analysis of selected gender-associated molecules in Schistosoma japonicum. Parasitology 135(5):575-83. Assay: Lactate dehydrogenase in worm.

To find more recent publications, please click here.

If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.