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.
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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
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.
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