Anti-HIF-2-alpha antibody [OTI2G5]

• WB

Supplier Data

Western blot - Anti-HIF-2-alpha antibody [OTI2G5] (AB157249)

False colour image of Western blot : Anti-HIF-2-alpha antibody [OTI2G5] staining at 1/500 dilution, shown in black; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab157249 was shown to bind specifically to HIF-2-alpha. A band was observed at 100 kDa in treated wild-type A549 cell lysates with no signal observed at this size in EPAS1 knockout cell line ab259774 (knockout cell lysate ab259779). To generate this image, wild-type and EPAS1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % BSA in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 20 seconds exposure time. Secondary antibodies used were HRP conjugated Goat anti-Mouse (H+L) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

All lanes:

Western blot - Anti-HIF-2-alpha antibody [OTI2G5] (ab157249) at 1/500 dilution

Lane 1:

Wild-type A549 Untreated (DFO Control) cell lysate at 20 µg

Lane 2:

Wild-type A549 Treated DFO (1 mM, 24 h) cell lysate at 20 µg

Lane 2:

Western blot - Human EPAS1 (HIF-2-alpha) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-epas1-hif-2-alpha-knockout-a549-cell-line-ab259774'>ab259774</a>)

Lane 3:

EPAS1 knockout A549 Untreated (DFO Control) cell lysate at 20 µg

Lane 4:

EPAS1 knockout A549 Treated DFO (1 mM, 24 h) cell lysate at 20 µg

Predicted band size: 96 kDa

Observed band size: 100 kDa

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• WB

Unknown

Western blot - Anti-HIF-2-alpha antibody [OTI2G5] (AB157249)

HEK293T cell lysates were generated from transient transfection of the cDNA clone (RC216194)

All lanes:

Western blot - Anti-HIF-2-alpha antibody [OTI2G5] (ab157249) at 1/2000 dilution

Lane 1:

HEK293T cells transfected with pCMV6-ENTRY control at 5 µg

Lane 2:

HEK293T cells transfected with pCMV6-ENTRY HIF2 alpha at 5 µg

Predicted band size: 96 kDa

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