Anti-NF-kB p65 antibody [E379] - Mouse IgG2a (Chimeric) - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NF-kB p65 antibody [E379] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307841)

This data was developed using ab307840, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling NF-kB p65 with ab307840 at 1/1000 (0.881 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human tonsil. The section was incubated with ab307840 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NF-kB p65 antibody [E379] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307841)

This data was developed using ab307840, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling NF-kB p65 with ab307840 at 1/1000 (0.881 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human ovarian cancer. The section was incubated with ab307840 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• WB

Supplier Data

Western blot - Anti-NF-kB p65 antibody [E379] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307841)

This data was developed using ab307840, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : In Western blot, ab307840 was shown to bind specifically to NF-kB p65. A band was observed at 65 kDa in wild-type HAP1 cell lysates with whereas no signal observed at this size in NF-kB p65 knockout cell lysate. The samples were run on a Bis-Tris gel. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time :

All lanes:

Western blot - Anti-NF-kB p65 antibody [E379] - Mouse IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/nf-kb-p65-antibody-e379-mouse-igg2a-chimeric-ab307840'>ab307840</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell line) cell lysate at 20 µg

Lane 2:

NF-kB p65 knockout HAP1 cell lysate at 20 µg

Lane 3:

HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) (ZB-2305) at 1/10000 dilution

Observed band size: 65 kDa

false

Exposure time: 10s