Anti-PDHA1 antibody [9H9AF5]

• WB

Lab

Western blot - Anti-PDHA1 antibody [9H9AF5] (AB110330)

Lanes 1 - 3 : Merged signal (red and green). Green - ab110330 observed at 43 kDa. Red - loading control, ab52866, observed at 50 kDa.

ab110330 was shown to specifically react with PDHA1 in wild-type HeLa cells as signal was lost in PDHA1 knockout cells. Wild-type and PDHA1 knockout samples were subjected to SDS-PAGE. ab110330 and ab52866 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/1000 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:

Western blot - Anti-PDHA1 antibody [9H9AF5] (ab110330) at 1 µg/mL

Lane 1:

Wild-type HeLa whole cell lysate at 20 µg

Lane 2:

PDHA1 knockout HeLa whole cell lysate at 20 µg

Lane 3:

HEK-293 whole cell lysate at 20 µg

Predicted band size: 43 kDa

Observed band size: 43 kDa

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• Flow Cyt

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Flow Cytometry - Anti-PDHA1 antibody [9H9AF5] (AB110330)

Flow cytometric analysis using ab110330 at 1μg/ml staining PDHA1 in HL60 cells (blue). Isotype control antibody (red).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PDHA1 antibody [9H9AF5] (AB110330)

Immunocytochemistry analysis using ab110330 at 5μg/ml staining PDHA1 in cultured, normal Human embryonic lung fibroblasts (strain MRC5) followed by AlexaFluor® 488 goat anti-mouse IgG1 secondary antibody (2 μg/ml).

• ICC/IF

AbReview39993****

Immunocytochemistry/ Immunofluorescence - Anti-PDHA1 antibody [9H9AF5] (AB110330)

Immunocytochemical analysis of SH-SY5Y human neuroblastoma labeling PDHA1 with ab110330 at 1/500 dilution.

'This image is courtesy of an Abreview submitted by George Allen'

• WB

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Western blot - Anti-PDHA1 antibody [9H9AF5] (AB110330)

All lanes:

Western blot - Anti-PDHA1 antibody [9H9AF5] (ab110330) at 1 µg/mL

Lane 1:

Isolated mitochondria from Human heart at 10 µg

Lane 2:

Isolated mitochondria from Bovine heart at 4 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

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• WB

CiteAb

Western blot - Anti-PDHA1 antibody [9H9AF5] (AB110330)

PDHA1 western blot using anti-PDHA1 antibody [9H9AF5] ab110330. Publication image and figure legend from Zhang, Y., Zhao, Z., et al., 2016, PLoS One, PubMed 26930489.

ab110330 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab110330 please see the product overview.

Induction of PDH phosphorylation by ATP.A. PDH phosphorylation. Mitochondria lysate was incubated with ATP at multiple dosages for 30 minutes at 37°C. PDH phosphorylation was quantified in Western blot and the results are expressed in bar figure B. Protein levels of PDKs. The proteins of PDK2 and PDK4 were determined in Western blot and the signals were quantified. The experiments were performed three times with consistent results, and the representative blot is shown. In the bar figure, the result represents mean ± SE (n = 3). * p<0.05 compared with the control.

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