Anti-HIF-2-alpha antibody [EPR19656]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] (AB207607)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, untreated or treated with deferoxamine mesylate salt (1mM, 24h), labeling HIF-2-alpha with ab207607 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on HeLa cells treated with deferoxamine mesylate salt (1mM, 24h).

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IP

Unknown

Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] (AB207607)

HIF-2-alpha was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell treated with 1mM DFO for 24h, cell lysate 10 μg with ab207607 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab207607 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate (10 μg) treated with 1mM DFO for 24h

Lane 2 : ab207607 IP in DFO treated HeLa cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab207607 in HeLa cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 75 seconds

All lanes:

Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

Predicted band size: 96 kDa

false

• WB

Supplier Data

Western blot - Anti-HIF-2-alpha antibody [EPR19656] (AB207607)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with the literature, and the expression level can be induced under hypoxia or treated with hypoxia mimetic chemicals such as DFO (PMID : 25587023, 10934146, 23958427 and 21571835).

All lanes:

Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607) at 1/1000 dilution

Lane 1:

Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

Whole cell lysate of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 1mM deferoxamine (DFO) mesylate salt for 24h at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 96 kDa

Observed band size: 110 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-HIF-2-alpha antibody [EPR19656] (AB207607)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with the literature, and the expression level can be induced under hypoxia or treated with hypoxia mimetic chemicals such as CoCl₂ (PMID : 21571835, 10934146 and 23958427).

All lanes:

Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607) at 1/1000 dilution

Lane 1:

Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

Whole cell lysate of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 100μM CoCl2 for 24h at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 96 kDa

Observed band size: 110 kDa

false

Exposure time: 3min