Human LDHC (L-lactate dehydrogenase C chain) ELISA Kit (DEIA-FN796)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
96T
Sample
serum, plasma, cell culture supernatants, tissue homogenate
Species Reactivity
Human
Intended Use
For quantitative detection of Human LDHC (L-lactate dehydrogenase C chain) in serum, plasma, tissue homogenates and other biological fluids.
Contents of Kit
1. 96-well strip plate (Dismountable), 1 plate
2. Lyophilized Standard, 2 vials
3. Sample/Standard dilution buffer, 20 mL
4. Biotin-detection antibody (Concentrated), 120 uL
5. Antibody dilution buffer, 10 mL
6. HRP-Streptavidin Conjugate(SABC), 120 uL
7. SABC dilution buffer, 10 mL
8. TMB substrate, 10 mL
9. Stop solution, 10 mL
10. Wash buffer (25X), 30 mL
11. Plate Sealer, 5 pieces
12. Product Manual, 1 copy
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Precision
Intra-Assay: CV<8%
Inter-Assay: CV<10%
Detection Range
1.56-100 ng/mL
Sensitivity
0.938 ng/mL
Standard Curve

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References


Human testis-specific lactate dehydrogenase-C promoter drives overexpression of mouse lactate dehydrogenase-1 cDNA in testes of transgenic mice

JOURNAL OF EXPERIMENTAL ZOOLOGY

Authors: Markert, CL; Amet, TM; Goldberg, E

The three isozymes of lactate dehydrogenase, each encoded by a separate gene, are developmentally regulated and differentially expressed in tissue-specific patterns. The lactate dehydrogenase-C (LDHC, mouse Ldh3) gene is temporally expressed exclusively in the germ line during spermatogenesis, whereas lactate dehydrogenase-A (LDHA, mouse Ldh1) and B (LDHB, mouse Ldh2) genes are active in somatic tissues. To determine, therefore, whether overexpression of Ldh1 would perturb spermatogenesis, we constructed a transgene in which a sequence from the promoter region of human LDHC was coupled with mouse Ldh1 cDNA. Among nine (three males, six females) founder lines that were identified as being transgenic for the construct, one male transmitted the gene through its germ line. Homo- and heterotetramers containing the LDH-A subunit were detected in homogenates of testes from transgenic animals. We conclude that the human LDHC promoter contains the necessary regulatory sequence(s) for specific expression of mouse Ldh1 as a transgene during spermatogenesis. The fertility of the founder animal was not impaired. J. Exp. Zool. 282:171-178, 1998. (C) 1998 Wiley-Liss, Inc.

Heat-induced apoptosis of mouse meiotic cells is suppressed by ectopic expression of testis-specific calpastatin

JOURNAL OF ANDROLOGY

Authors: Somwaru, L; Li, SM; Doglio, L; Goldberg, E; Zirkin, BR

Calpastatin is a naturally occurring inhibitor of calpain, a protease involved in apoptotic cell death. A testis-specific isoform of calpastatin (tCAST) has been identified that is transcribed in haploid germ cells but not in spermatocytes. To investigate the possible function(s) of tCAST, we tested the hypothesis that the ectopic expression of calpastatin in spermatocytes would suppress the death of these cells in response to an apoptosis-inducing stimulus in vivo. To this end, the 5'-flanking region of the mouse Idhc gene was linked to tCAST, and transgenic mice were generated. Immunohistochemical analysis revealed that, in contrast to control sections in which the signal for tCAST was seen in round spermatids, intense staining was visualized in pachytene spermatocytes in the transgenic animals, indicating that the strategy we used to generate the transgenic animals resulted in the ectopic expression of tCAST in spermatocytes. We then tested the effect of a short period of heating on germ cell apoptosis in the testes of wild-type and transgenic mice. Pachytene spermatocytes were the major germ cell type seen to undergo apoptosis after heat treatment. There were no differences in the number of apoptotic germ cells per seminiferous tubule between wild-type and tCAST transgenic control mice; thus, there was no apparent effect of the transgene on normal apoptosis. Heating resulted in increased numbers of TUNEL-positive germ cells in both wild-type and tCAST transgenic mice, as well as increased testicular DNA fragmentation. Heating the tCAST transgenic mouse testes resulted in significantly fewer apoptotic cells per seminiferous tubule than in wild-type mice at both 8 and 24 hours after treatment. Thus, as hypothesized, the ectopic expression of tCAST in pachytene spermatocytes suppressed germ cell apoptosis.

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