All lanes : Anti-PDHA1 antibody [8D10E6] (ab110334) at 0.5 µg/ml

Lane 1 : Wild-type HeLa whole cell lysate
Lane 2 : PDHA1 knockout HeLa whole cell lysate
Lane 3 : HEK-293 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 43 kDa
Observed band size: 43 kDa

Lanes 1 - 3: Merged signal (red and green). Green - ab110334 observed at 43 kDa. Red - loading control, ab52866, observed at 50 kDa.

ab110334 was shown to recognize PDHA1 in wild-type HeLa cells as signal was lost at the expected MW in PDHA1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PDHA1 knockout samples were subjected to SDS-PAGE. Ab110334 and ab52866 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 0.5 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes : Anti-PDHA1 antibody [8D10E6] (ab110334) at 1 µg/ml

Lane 1 : Isolated mitochondria from human heart at 5 µg
Lane 2 : Isolated mitochondria from bovine heart at 1 µg
Lane 3 : Isolated mitochondria from rat heart at 10 µg
Lane 4 : Isolated mitochondria from mouse heart at 10 µg
Lane 5 : HepG2 cell lysate at 20 µg

Predicted band size: 43 kDa

Immunocytochemistry analysis using ab110334 at 1 µg/ml staining PDHA1 in HeLa (human epithelial cell line from cervix adenocarcinoma) cells (4% paraformaldehyde fixed and Triton X-100 permeabilized).
The secondary antibody was (green) Alexa Fluor^® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution. DAPI was used to stain the cell nuclei (blue).

Flow cytometric analysis using ab110334 at 1 µg/ml staining PDHA1 in HL-60 (human promyelocytic leukemia cell line) cells (blue). Isotype control antibody (red).