Anti-HIF1 beta antibody [29/HIF-1b] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [29/HIF-1b] - BSA and Azide free (AB289560)

This data was developed using ab289550 the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling HIF1 beta with ab289550 at 1/50 (21.24 μg/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line ab179513. Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/500 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab289550 at 1/50 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/500 dilution.

Negative control 2 : ab179513 Anti-beta Tubulin rabbit monoclonal antibody at 1/200 dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [29/HIF-1b] - BSA and Azide free (AB289560)

This data was developed using ab289550 the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling HIF1 beta with ab289550 at 1/50 (21.24 μg/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line ab179513. Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/500 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab289550 at 1/50 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/500 dilution.

Negative control 2 : ab179513 Anti-beta Tubulin rabbit monoclonal antibody at 1/200 dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-HIF1 beta antibody [29/HIF-1b] - BSA and Azide free (AB289560)

This data was developed using ab289550, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer : 5% NFDM/TBST.

Lysates should be made freshly and used in WB immediately to minimize protein degradation.

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-HIF1 beta antibody [29/HIF-1b] (<a href='/en-us/products/primary-antibodies/hif1-beta-antibody-29-hif-1b-ab289550'>ab289550</a>) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 87 kDa

Observed band size: 95 kDa

false