Human CREB Antibody

Detection of Human CREB by Western Blot.

Western blot shows lysates of HT-29 human colon adenocarcinoma cell line, HepG2 human hepatocellular carcinoma cell line, MBA-MB-468 human breast cancer cell line, and MCF-7 human breast cancer cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human CREB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2989) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for CREB at approximately 43 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of CREB-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 for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. CREB/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human CREB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2989) 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. Thefospromoter was detected by standard PCR.

CREB in Human Breast.

CREB was detected in immersion fixed paraffin-embedded sections of human breast using 10 µg/mL Goat Anti-Human CREB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2989) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the nucleus in stromal cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.