Human/Mouse c-Rel Antibody

Detection of Human/Mouse c-Rel by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line, A20 mouse B cell lymphoma cell line, and CH-1 mouse B cell lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse c-Rel Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2699) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for c-Rel at approximately 69 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of c‑Rel-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL and 200 ng/mL PMA and calcium ionomycin for overnight was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. c-Rel/DNA complexes were immuno-precipitated using 5 µg Goat Anti-Human/Mouse c-Rel Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2699) 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. Thep21promoter was detected by standard PCR.

c-Rel in THP-1 Human Cell Line. c-Rel was detected in immersion fixed THP-1 human acute monocytic leukemia cell line treated with LPS using Goat Anti-Human/Mouse c-Rel Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2699) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell nuclei upon stimulation with LPS. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Mouse c‑Rel by Simple Western^TM. Simple Western lane view shows lysates of CH-1 mouse B cell lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for c-Rel at approximately 71 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse c-Rel Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2699) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.