Anti-NF-kB p65 抗体 [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

Lab

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

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

Western blot : Anti-NF-kB p65 antibody [EP2161Y] ab76311 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 60 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in RELA knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk 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 then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (<a href='/products/primary-antibodies/nf-kb-p65-antibody-ep2161y-ab76311'>ab76311</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 cell lysates at 20 µg

Lane 2:

Western blot - Human RELA knockout MCF7 cell line (ab287740) at 40 µg

Lane 3:

PANC-1 cell lysates at 20 µg

Lane 4:

Saos-2 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 60 kDa

Observed band size: 60 kDa

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