|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.|
Chromatin Immunoprecipitation (ChIP)
|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 immunoprecipitated 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. The p21 promoter was detected by standard PCR.|
Detection of Mouse c‑Rel by Simple WesternTM. 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.
c-Rel/Rel, the cellular counterpart of the v-Rel oncogene of the avian reticuloendotheliosis retrovirus, is a 69 kDa class II member of the Rel/NK-kappa B family of transcription factors. All Rel family members contain a RHD (Rel homology domain) that is involved in dimerization, DNA and I kappa B binding, and nuclear localization. Class II members contain an additional C-terminal transcriptional activation segment. Rel both homodimerizes and heterodimerizes with multiple family members. Following dimerization, Rel complexes initiate transcription by acting on decameric DNA motifs termed kappa B binding sites. The important role of c-Rel in B-cell development, growth, and survival is well documented. c-Rel is also involved in responses to auto-antigens, allo-antigens, allergens and pathogens and may contribute to the development of certain human lymphoid cell cancers.
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