Cell Signaling Technology

Product Pathways - Metabolism

Phospho-LDHA (Tyr10) Antibody #8176

Applications Reactivity Sensitivity MW (kDa) Source
W H M R Endogenous 37 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-LDHA (Tyr10) Antibody recognizes endogenous levels of LDHA protein only when phosphorylated at Tyr10.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr10 of human LDHA protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Phospho-LDHA (Tyr10) Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells transfected with a construct expressing DYKDDDDK-tagged wild-type (WT) full-length human LDHA (WT hLDHA-DDK) alone or in conjuction with full-length human FGFR1 (hFGFR1), using Phospho-LDHA (Tyr10) Antibody (upper) or DYKDDDDK Tag (9A3) Mouse mAb #8146 (lower). The DYKDDDDK-tagged WT hLDHA expression construct was kindly provided by Dr. Jing Chen at Emory University School of Medicine.

Background

Lactate dehydrogenase (LDH) catalyzes the interconversion of pyruvate and NADH to lactate and NAD+. When the oxygen supply is too low for mitochondrial ATP production, this reaction recycles NADH generated in glycolysis to NAD+, which reenters glycolysis. The major form of LDH found in muscle cells is the A (LDHA) isozyme. The LDHA promoter contains HIF-1α binding sites (1). LDHA expression is induced under hypoxic conditions (2). During intensive and prolonged muscle exercise, lactate accumulates in muscle cells when the supply of oxygen does not meet demand. When oxygen levels return to normal, LDH converts lactate to pyruvate to generate ATP in the mitochondrial electron transport chain.

Recent studies indicate that LDHA is phosphorylated at Tyr10 and Tyr83 by the receptor tyrosine kinase FGFR1 (3). Phosphorylation at Tyr10 up-regulates the activity of LDHA and is found in a variety of human cancer cells (3). Results further suggest that tyrosine phosphorylation regulates the NADH/NAD+ redox homeostasis and thus promotes the Warburg effect and tumor cell growth (3).

  1. Semenza, G.L. et al. (1996) J Biol Chem 271, 32529-37.
  2. Semenza, G.L. (2007) Biochem J 405, 1-9.
  3. Fan, J. et al. (2011) Mol Cell Biol 31, 4938-50.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Products