Validated using a knockout cell line

Anti-CREB antibody [E306] (ab32515)


  • Product name
    Anti-CREB antibody [E306]
    See all CREB primary antibodies
  • Description
    Rabbit monoclonal [E306] to CREB
  • Host species
  • Specificity
    This antibody recognizes CREB, and can also recognize the splice isoform of CREB.
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-P, IPmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide within Human CREB aa 250-350. The exact sequence is proprietary.

  • Positive control
    • ICC/IF: PC 12 cells. WB: A431 cell lysate. IHC-P: Prostatic carcinoma tissue.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.



Our Abpromise guarantee covers the use of ab32515 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/1000. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa).
ICC/IF 1/250 - 1/500.
IHC-P Use at an assay dependent concentration.
IP 1/80.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Function
      This protein binds the cAMP response element (CRE), a sequence present in many viral and cellular promoters. CREB stimulates transcription on binding to the CRE. Transcription activation is enhanced by the TORC coactivators which act independently of Ser-133 phosphorylation. Implicated in synchronization of circadian rhythmicity.
    • Involvement in disease
      Defects in CREB1 may be a cause of angiomatoid fibrous histiocytoma (AFH) [MIM:612160]. A distinct variant of malignant fibrous histiocytoma that typically occurs in children and adolescents and is manifest by nodular subcutaneous growth. Characteristic microscopic features include lobulated sheets of histiocyte-like cells intimately associated with areas of hemorrhage and cystic pseudovascular spaces, as well as a striking cuffing of inflammatory cells, mimicking a lymph node metastasis. Note=A chromosomal aberration involving CREB1 is found in a patient with angiomatoid fibrous histiocytoma. Translocation t(2;22)(q33;q12) with CREB1 generates a EWSR1/CREB1 fusion gene that is most common genetic abnormality in this tumor type.
    • Sequence similarities
      Belongs to the bZIP family.
      Contains 1 bZIP domain.
      Contains 1 KID (kinase-inducible) domain.
    • Post-translational
      Stimulated by phosphorylation. Phosphorylation of both Ser-133 and Ser-142 in the SCN regulates the activity of CREB and participates in circadian rhythm generation. Phosphorylation of Ser-133 allows CREBBP binding (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
      Sumoylated by SUMO1. Sumoylation on Lys-304, but not on Lys-285, is required for nuclear localization of this protein. Sumoylation is enhanced under hypoxia, promoting nuclear localization and stabilization.
    • Cellular localization
    • Information by UniProt
    • Database links
    • Alternative names
      • Active transcription factor CREB antibody
      • cAMP response element binding protein 1 antibody
      • cAMP response element binding protein antibody
      • cAMP responsive element binding protein 1 antibody
      • cAMP-responsive element-binding protein 1 antibody
      • CREB antibody
      • CREB-1 antibody
      • CREB1 antibody
      • CREB1_HUMAN antibody
      • Cyclic AMP-responsive element-binding protein 1 antibody
      • MGC9284 antibody
      • OTTHUMP00000163864 antibody
      • OTTHUMP00000163865 antibody
      • OTTHUMP00000206660 antibody
      • OTTHUMP00000206662 antibody
      • OTTHUMP00000206667 antibody
      • Transactivator protein antibody
      see all


    • All lanes : Anti-CREB antibody [E306] (ab32515) at 0.03 µg/ml

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
      Lane 2 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg
      Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
      Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 37 kDa
      Observed band size: 40 kDa (why is the actual band size different from the predicted?)

      Exposure time: 15 seconds

      Blocking and diluting buffer: 5% NFDM/TBST.

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: CREB knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: A431 cell lysate(20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab32515 observed at 44 kDa. Red - loading control, ab18058, observed at 214 kDa.

      ab32515 was shown to recognize CREB in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when CREB knockout samples examined. Wild-type and knockout samples were subjected to SDS-PAGE. ab32515 and ab18058 (loading control to Vinculin) diluted to 1/500 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • ab32515 staining CREB in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. ab150081, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counterstained with DAPI.

      See Abreview

    • Ab32515, at a 1/500 dilution, staining CREB in paraffin embedded prostatic carcinoma tissue sections by Immunohistochemistry.


    This product has been referenced in:
    • Wu XP  et al. MicroRNA-365 alleviates morphine analgesic tolerance via the inactivation of the ERK/CREB signaling pathway by negatively targeting ß-arrestin2. J Biomed Sci 25:10 (2018). WB . Read more (PubMed: 29415719) »
    • Li X  et al. Cisplatin Enhances Hepatitis B Virus Replication and PGC-1a Expression through Endoplasmic Reticulum Stress. Sci Rep 8:3496 (2018). Read more (PubMed: 29472690) »

    See all 27 Publications for this product

    Customer reviews and Q&As

    Western blot
    Zebrafish Tissue lysate - whole (Testicle)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    5 µg
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Apr 17 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Western blot
    Human Cell lysate - whole cell (Hela)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    10 µg
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

    Dr. Lin Zhu

    Verified customer

    Submitted Sep 04 2017

    Immunocytochemistry/ Immunofluorescence
    Human Cell (HeLa)
    Yes - 0.5% Triton-X100 in PBS

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Nov 05 2014

    Thank you for this additional information. I would recommend testing under reducing and denaturing conditions rather than a native gel, as this is how we have tested this product. Otherwise your protocol seems good. Please let me know if you are still ...

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    Thank you for confirming this information and for your help and cooperation with this case.

    As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order...

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    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    I appreciate the time you have spent in the laboratory and understand y...

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    Thank you for contacting Abcam.

    The concentration for ab32515, lot #GR50483-2, is**********.

    Please let me know if there is anything else I can help you with.

    Codes offre de test :
    ab32515 : *****
    ab32096 : *****
    ab7217 : *****
    ab12135 : *****
    ab1791 : *****

    Date d’expiration : JJ MM AAAA

    Je suis ravi que vous souhaitiez participer ànotre offre et t...

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