|Detection of Human NF kappa B1 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse NF kappa B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2697) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for NF kappa B1 at approximately 50 and 105 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Chromatin Immunoprecipitation (ChIP)
|Detection of NF kappa B1-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin overnight was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. NF kappa B1/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse NF kappa B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2697) or control antibody (Catalog # AB‑108‑C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The bcl-2 promoter was detected by standard PCR. This application has not been tested in mouse samples.|
Nuclear Factor kappa B1 (NF kappa B1 or NF kappa B p50) is a member of the rel/NF kappa B family of transcription factors. NF kappa B1 dimerizes with other members of the rel/NF kappa B family to regulate expression of genes that participate in immune, apoptotic, and oncogenic processes.