Human/Mouse NFkB1 Antibody

Detection of Human NF kappaB1 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.

Detection of NF kappaB1-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. Thebcl-2promoter was detected by standard PCR. This application has not been tested in mouse samples.