Human, Zebrafish Predicted to work with:
Mouse, Rat, Rabbit, Chimpanzee, Chinese hamster
Synthetic peptide corresponding to Human Lactate Dehydrogenase aa 300 to the C-terminus conjugated to Keyhole Limpet Haemocyanin (KLH). (Peptide available as ab47009, ab47009)
This antibody gave a positive signal in the following whole cell lysates:
HeLa (Human epithelial carcinoma cell line)
Jurkat (Human T cell lymphoblast-like cell line)
A431 (Human epithelial carcinoma cell line)
HEK293 (Human embryonic kidney cell line)
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
IHC image of Lactate Dehydrogenase staining in human breast carcinoma FFPE section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab47010, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
ICC/IF image of ab47010 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab47010, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
LDHA was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to LDHA and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab47010.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody (ab47010)This image is from an anonymous abreview.
ab47010 at a 1/400 dilution staining LDHA in human skeletal muscle sections by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections) incubated for 20 minutes at 25°C. Heat mediated antigen retrieval used, EDTA ph 9.0, 100°C for 20 minutes. Secondary used undiluted, goat anti-mouse/rabbit IgG conjugated to HRP polymer.
Gaude E et al. NADH Shuttling Couples Cytosolic Reductive Carboxylation of Glutamine with Glycolysis in Cells with Mitochondrial Dysfunction. Mol Cell69:581-593.e7 (2018).
Read more (PubMed: 29452638) »