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.

CREB in Human Dorsal Root Ganglion. CREB was detected in immersion fixed paraffin-embedded sections of human dorsal root ganglion using Goat Anti-Human CREB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2989) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

CREB in MCF‑7 Human Cell Line. CREB was detected in immersion fixed MCF-7 human breast cancer cell line using Goat Anti-Human CREB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2989) 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 nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

CREB in MCF‑7 Human Cell Line. CREB was detected in immersion fixed MCF-7 human breast cancer cell line using Goat Anti-Human CREB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2989) at 0.5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.