Anti-LDHA antibody (ab47010)

Western blot - LDHA antibody (ab47010)

All lanes : LDHA antibody (ab47010) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 37 kDa
Observed band size : 36 kDa (why is the actual band size different from the predicted?)

Immunocytochemistry/ Immunofluorescence - LDHA antibody (ab47010)

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).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - LDHA antibody (ab47010)

IHC image of Lactate Dehydrogenase staining in human breast carcinoma FFPE section, performed on a Leica Bond^TM 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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - LDHA antibody (ab47010)

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.

This image is from an anonymous abreview.

Western blot - Anti-LDHA antibody (ab47010)

Anti-LDHA antibody (ab47010) at 1 µg/ml + LDHA protein (His tag) (ab93699) at 0.1 µg

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Exposure time : 10 seconds