All lanes : Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333) at 1 µg/ml

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : DLAT knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HL-60 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 54 , 69 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab110333 observed at 72 kDa. Red - loading control, ab181602, observed at 38 kDa.

ab110333 was shown to specifically react with in wild-type HAP1 cells as signal was lost in DLAT knockout cells. Wild-type and DLAT knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab110333 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes : Anti-Pyruvate dehydrogenase E2/E3bp antibody [13G2AE2BH5] (ab110333) 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: 54 , 69 kDa

ab110333 staining Pyruvate dehydrogenase E2/E3bp in Human HUVEC by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 pH 7.4 for 5 minutes and blocked with 5% BSA for 20 minutes at room temperature. Samples were incubated with primary antibody (1/1000 in PBS) for 1 hour. A CF568-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.

See Abreview

Immunocytochemistry analysis using ab110333 at 1µg/ml staining Pyruvate dehydrogenase E2/E3bp in cultured, normal Human embryonic lung fibroblasts and an AlexaFluor® 488 goat anti-mouse IgG2a secondary antibody (2 ug/ml).

Immunohistological analysis using ab110333 at 1/1000 dilution staining Pyruvate dehydrogenase E2/E3bp in Human cerebellum tissue (Formalin-fixed, Paraffin-embedded).
Note: Immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells.

Flow cytometric analysis using ab110333 at 1µg/ml staining Pyruvate dehydrogenase E2/E3b in HL60 cells (blue). Isotype control antibody (red).