Overview

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

    Synthetic peptide (Human), corresponding to residues before DNA binding motif of human CREB.

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

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

    Alternative versions available:

    Anti-CREB antibody (Alexa Fluor® 647) [E306] (ab194312)

    Anti-CREB antibody (HRP) [E306] (ab194313)

Properties

Applications

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.
Flow Cyt 1/40.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/80.

Target

  • FunctionThis 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 diseaseDefects 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 similaritiesBelongs to the bZIP family.
    Contains 1 bZIP domain.
    Contains 1 KID (kinase-inducible) domain.
  • Post-translational
    modifications
    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 localizationNucleus.
  • 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

Anti-CREB antibody [E306] images



  • Predicted band size : 37 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ProteinX 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 when CREB knockout samples were used, along with additional cross-reactive bands. Wild-type and CREB 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) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 h 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.
  • Overlay histogram showing HEK293 cells stained with ab32515 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32515, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Anti-CREB antibody [E306] (ab32515) at 1/1000 dilution + A431 cell lysate

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

References for Anti-CREB antibody [E306] (ab32515)

This product has been referenced in:
  • Pan W  et al. Effects of dihydrotestosterone on synaptic plasticity of the hippocampus in mild cognitive impairment male SAMP8 mice. Exp Ther Med 12:1455-1463 (2016). IHC-P ; Mouse . Read more (PubMed: 27588067) »
  • Snow WM  et al. Morris Water Maze Training in Mice Elevates Hippocampal Levels of Transcription Factors Nuclear Factor (Erythroid-derived 2)-like 2 and Nuclear Factor Kappa B p65. Front Mol Neurosci 8:70 (2015). WB ; Mouse . Read more (PubMed: 26635523) »

See all 12 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.5% Triton-X100 in PBS
Fixative Paraformaldehyde
Username

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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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

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