CREB Antibody [Unconjugated]

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.

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.

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. The fos promoter was detected by standard PCR.

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 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 at4 °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 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.

• Reactivity: Hu
• Applications: WB, IHC, ChIP, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CREB Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human CREB
  Lys136-Asp341
  Accession # P16220
• Specificity: Detects human CREB in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CREB1

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Chromatin Immunoprecipitation (ChIP) 5 ug/5 x 10^6 cells
  • Immunocytochemistry 5-15 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CREB Antibody [Unconjugated]

• active transcription factor CREB
• cAMP responsive element binding protein 1
• cAMP-response element-binding protein-1
• cAMP-responsive element-binding protein 1
• CREB
• CREB1
• CREB-1
• cyclic AMP-responsive element-binding protein 1
• MGC9284
• transactivator protein

Background

The cAMP response element binding protein (CREB), also known as CREB1, belongs to the bZIP superfamily of transcription factors, containing a basic domain that mediates DNA binding and a leucine zipper domain that facilitates dimerization. Within the promoter of target genes, CREB dimers bind cAMP response elements, defined by the palindromic consensus sequence TGACGTCA.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CREB Antibody (AF2989)(1)

Publication using AF2989

• MX Liu, M Gao, CZ Li, CZ Yu, H Yan, C Peng, Y Li, CG Li, ZL Ma, Y Zhao, MF Pu, LL Miao, XM Qi, J Ren Dicer1/miR-29/HMGCR axis contributes to hepatic free cholesterol accumulation in mouse non-alcoholic steatohepatitis Acta Pharmacol. Sin, 2017;0(0):. 2017 [PMID: 28112179] (Western Blot, Mouse)

FAQs for CREB Antibody (AF2989). (Showing 1 - 1 of 1 FAQ).

1. May I know which creb1 antibody is good for ChIP? I found a paper using your rabbit polyclonal creb1 antibody for ChIP but they didn't indicate the catalog number.
   • We are not aware of any of our Creb1 antibodies being used in ChIP at this time. If you can provide us with the PMID of the paper you are referring to I can contact the author. Otherwise, you can use our Innovators Reward Program to try an antibody in a novel species and application.

Bioinformatics

• Gene Symbol: CREB1
• Uniprot:
  • P16220()