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Members of the rel/NFkappaB family of transcription factors are involved in the regulation of cellular responses, such as growth, development, and the inflammatory response. They share a structural motif known as the rel homology region (RHR), the C-terminal one third of which mediates protein dimerization. Complexes of p50 (NF-kB1) or p52 (NF-kB2) are generated through the processing of p105 and p100 precursors, respectively. These are usually associated with members of the Rel family (p65, c-Rel, Rel B). The homo- and heterodimer formed through combinations of NF-kB/Rel proteins bind distinct kB sites to regulate the transcription of different genes.
Our Abpromise guarantee covers the use of ab7970 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent dilution.|
|ICC||Use at an assay dependent concentration.|
|ChIP/Chip||Use at an assay dependent concentration.|
|EMSA||Use a concentration of 1 µg/ml.|
|IP||Use at an assay dependent dilution.|
|IHC-P||1/1000 - 1/4000. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.|
|WB||1/400 - 1/1000. Detects a band of approximately 64 kDa (predicted molecular weight: 69 kDa).Can be blocked with NF-kB p65 peptide (ab8005).|
|IHC-Fr||1/100. PubMed: 17003500|
|ICC/IF||Use at an assay dependent dilution. PubMed: 16818736|
|Flow Cyt||Use at an assay dependent concentration.|
ab7970 staining NF-kB p65 in Human AGS cells by ICC/IF (Immunocytochemistry/immunoflurescence). Cells were fixed with formaldehyde, permeabilzed with 0.025% Trton X-100 in TBS and blocked with 5% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/100) for 1 hour at 25°C. An Alexa Fluor®568-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
Western blotting starting dilution: 1/400-1/1000.
This image is a courtesy of Anonymous Abreview
Immunoperoxidase staining of formalin-fixed, paraffin-embedded human DLBCL showing nuclear expression of NF-kB p65 in the tumor cells.
ab7970 staining NF-kB p65 in Human stomach tissue sections by IHC-Fr (Immunohistochemistry - Froxen sections). Tissue samples were fixed with acetone and blocked with 5% serum for 1 hour at 25°C. Samples were incubated with primary antibody 1/1000 in blocking buffer for 1 hour at 25°C. An undiluted HRP-conjugated Goat polyclonal to rabbit IgG was used as secondary antibody.
ab7970 staining NF-kB p65 in mouse lung tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent xylene fixation before heat mediated antigen retrieval in citrate and then blocking with 1.5% serum was performed for 30 minutes at 37°C. The primary antibody was used diluted 1/500 and incubated with sample for 16 hours at 4°C. A Biotin conjugated polyclonal to rabbit IgG was used undiluted as secondary antibody. ABC staining system was used.
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