Human/Rat Lactate Dehydrogenase A/LDHA Antibody Summary
Accession # P00338
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Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human and Rat Lactate Dehydrogenase A/LDHA by Western Blot. Western blot shows lysates of Daudi human Burkitt's lymphoma cell line, HepG2 human hepatocellular carcinoma cell line, MOLT-4 human acute lymphoblastic leukemia cell line, and rat skeletal muscle tissue. PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human/Rat Lactate Dehydrogenase A/LDHA Monoclonal Antibody (Catalog # MAB9158) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Lactate Dehydrogenase A/LDHA at approximately 34-36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Lactate Dehydrogenase A/LDHA in Human Pancreas. Lactate Dehydrogenase A/LDHA was detected in immersion fixed paraffin-embedded sections of human pancreas using Rabbit Anti-Human/Rat Lactate Dehydrogenase A/LDHA Monoclonal Antibody (Catalog # MAB9158) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to exocrine cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Lactate Dehydrogenase A/LDHA in PC‑3 Human Cell Line. Lactate Dehydrogenase A/LDHA was detected in immersion fixed PC-3 human prostate cancer cell line using Rabbit Anti-Human/Rat Lactate Dehydrogenase A/LDHA Monoclonal Antibody (Catalog # MAB9158) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Lactate Dehydrogenase A/LDHA
LDHA (lactate dehydrogenase A chain; also LDH-M and PIG-19) is a 34-36 kDa member of the LDH family of enzymes. It is a part of a cytoplasmic complex that is found principally in hepatocytes and skeletal muscle. The LDHA complex catalyzes the conversion of pyruvate to lactate, thereby generating NAD, the final step in anerobic glycolysis. This NAD is essential for the subsequent generation of ATP. Human LDHA is 332 amino acids (aa) in length. It contains an N-terminal coenzyme binding region, a central catalytic site, and at least nine utilized Lys acetylation and two Tyr phosphorylation sites. It also undergoes ISGylation where a 17 kDa product of the ISG15 gene is covalently attached to LDHA in a ubiquitin-like manner. LDHA forms tetramers composed of two dimers of varying composition. The tetramer may be homotetrameric (four monomers), or contain from one to three substitute LDHB monomers typically found in heart muscle. There are multiple splice variants. One possesses a five aa substitution for aa 237-332, a second contains a 45 aa substitution for aa 230-332, a third shows a deletion of aa 82-139, while a fourth utilizes an alternative start site 29 aa upstream of the standard site. Over aa 2-92, human LDHA shares 93% aa sequence identity with mouse LDHA.
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