Western blot analysis was performed on membrane enriched extracts (30 ug lysate) of HeLa (Lane 1), MCF7 (Lane 2), Hep G2 (Lane 3), HEK-293 (Lane 4), IMR-32 (Lane 5), MDA-MB-231 (Lane 6), A-431 (Lane 7), K-562 (Lane 8) and Jurkat (Lane 9).The blots were probed with LDHA Mouse monoclonal Antibody (Product# MA5-17247, 1 ug/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 ug/ml, 1:4000 dilution). A 37 kDa band corresponding to LDHA was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Recombinant human protein purified from E.coli (His-LDHA)|
|Storage buffer||HEPES with 0.15M NaCl, 0.01% BSA, 50% glycerol|
|Contains||0.03% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||0.5-2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
A suggested positive control for this product is HeLa cells.
Lactate dehydrogenase (LDH) is the enzyme in the glycolytic pathway that converts pyruvate to lactate with concomitant interconversion of NADH and NAD+. In mammals, the enzyme is encoded by three genes: LDHA (M or muscle form), LDHB (H or heart form), and LDHC (X or testis form). The three human LDHs have 84-89% sequence similarities and 69-75% amino acid identities. There are five different LDH isoenzymes based on the proportion of M and H chains existing in the LDH tetrameric structure. The LDHA is hypoxia inducible and its expression is directly controlled by the transcriptional activity of the hypoxia inducible factor 1a (HIF1a). LDHB is also known to be upregulated in many cancers.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.