Validated using a knockout cell line
Recombinant
RabMAb

Anti-CREB antibody [E306] (ab32515)

Overview

  • Product name
    Anti-CREB antibody [E306]
    See all CREB primary antibodies
  • Description
    Rabbit monoclonal [E306] to CREB
  • Specificity
    This antibody recognizes CREB, and can also recognize the splice isoform of CREB.
  • Tested applications
    Suitable 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.

     

    Alternative versions available:

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

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

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

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

  • 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
    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 localization
    Nucleus.
  • 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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:
  • Liu Y  et al. Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis. Respir Res 18:38 (2017). WB ; Rat . Read more (PubMed: 28222740) »
  • Wu X  et al. CUG-binding protein 1 regulates HSC activation and liver fibrogenesis. Nat Commun 7:13498 (2016). WB ; Human . Read more (PubMed: 27853137) »

See all 15 Publications for this product

Product Wall

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

Dr. Lin Zhu

Verified customer

Submitted Sep 04 2017

Application
Western blot
Sample
Rat Tissue lysate - whole (brain)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
15 µg
Specification
brain
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jun 19 2017

Application
Western blot
Sample
Rat Tissue lysate - whole (brain)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
15 µg
Specification
brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jun 12 2017

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**********.

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

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