Anti-NF-kB p65 antibody [EP2161Y] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 antibody [EP2161Y] - BSA and Azide free (AB246347)

Immunocytochemistry/Immunofluorescence analysis HT-29 (human colorectal adenocarcinoma) labelling NF-kB p65 with purified ab76311 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

Control : PBS only

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76311).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] - BSA and Azide free (AB246347)

This data was developed using ab76311, the same antibody clone in a different buffer formulation.

Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling NF-kB p65 with purified ab76311 at 1/1000 dilution (1 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] - BSA and Azide free (AB246347)

This data was developed using ab76311, the same antibody clone in a different buffer formulation.

Flow Cytometry analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labeling NF-kB p65 with purified ab76311 at 1/1000 dilution (1 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

Supplier Data

Western blot - Anti-NF-kB p65 antibody [EP2161Y] - BSA and Azide free (AB246347)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : NF-κB p65 knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : A431 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab76311 observed at 70 kDa. Red - ab8245 loading control, observed at 37 kDa.

ab76311 was shown to specifically react with NF-κB p65 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when NF-κB p65 knockout samples were used. Wild-type and NF-κB p65 knockout samples were subjected to SDS-PAGE. ab76311 (NF-κB p65) and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76311).

All lanes:

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (<a href='/en-us/products/primary-antibodies/nf-kb-p65-antibody-ep2161y-ab76311'>ab76311</a>)

Predicted band size: 60 kDa

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