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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CREB antibody [E306] (ab32515)

Reviews (3)Q&A (5)References (94)
Western blot - Anti-CREB antibody [E306] (ab32515)
  • Western blot - Anti-CREB antibody [E306] (ab32515)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB antibody [E306] (ab32515)
  • Anti-CREB antibody [E306] (ab32515)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E306] to CREB
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free HRP

Overview

  • Product name

    Anti-CREB antibody [E306]
    See all CREB primary antibodies

  • Description

    Rabbit monoclonal [E306] to CREB

  • Host species

    Rabbit

  • Specificity

    This antibody recognizes CREB, and can also recognize the splice isoform of CREB.

  • Tested applications

    Suitable for: WB, IHC-Pmore 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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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

The Abpromise guarantee

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 (2)
1/500 - 1/1000. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/500 - 1/1000. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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
    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

    see all

  • 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

Images

  • Western blot - Anti-CREB antibody [E306] (ab32515)
    Western blot - Anti-CREB antibody [E306] (ab32515)
    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

    Secondary
    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.

  • Western blot - Anti-CREB antibody [E306] (ab32515)
    Western blot - Anti-CREB antibody [E306] (ab32515)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB antibody [E306] (ab32515)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB antibody [E306] (ab32515)

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

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-CREB antibody [E306] (ab32515)
    Anti-CREB antibody [E306] (ab32515)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (94)

Publishing research using ab32515? Please let us know so that we can cite the reference in this datasheet.

ab32515 has been referenced in 94 publications.

  • Li J  et al. Prophylactic Melatonin Treatment Ameliorated Propofol-Induced Cognitive Dysfunction in Aged Rats. Neurotox Res 39:227-239 (2021). PubMed: 33159663
  • Song D  et al. GABAB receptor antagonist promotes hippocampal neurogenesis and facilitates cognitive function recovery following acute cerebral ischemia in mice. Stem Cell Res Ther 12:22 (2021). PubMed: 33413637
  • Zhang Y  et al. Dexmedetomidine attenuates sevoflurane-induced neurocognitive impairment through a2-adrenoceptors. Mol Med Rep 23:N/A (2021). PubMed: 33179100
  • Ling J  et al. Edaravone Improves Intermittent Hypoxia-Induced Cognitive Impairment and Hippocampal Damage in Rats. Biol Pharm Bull 43:1196-1201 (2020). PubMed: 32475934
  • Liu ZY  et al. MicroRNA-144 regulates angiotensin II-induced cardiac fibroblast activation by targeting CREB. Exp Ther Med 20:2113-2121 (2020). PubMed: 32765685
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-8 of 8 Abreviews or Q&A

Western blot abreview for Anti-CREB antibody [E306]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Zebrafish Tissue lysate - whole (Testicle)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
5 µg
Specification
Testicle
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 17 2018

Western blot abreview for Anti-CREB antibody [E306]

 Excellent
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Lin Zhu

Verified customer

Submitted Sep 04 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-CREB antibody [E306]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Permeabilization
Yes - 0.5% Triton-X100 in PBS
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Kirk Mcmanus

Verified customer

Submitted Nov 05 2014

Question

seeing a band at wrong MW
1) What species and sample type were you testing?

Mouse RAW267.4 cell line.


2) How much protein was loaded?

25 micrograms

3) What is your blocking step?

5% milk in TBST

4) What was the primary antibody dilution and incubation?

1:1000, 4oC, overnight.

5) WHat was the secondary dilution and incubation?

1:3000, room temp, 90mins

6) Was the gel run under reducing and denaturing conditions?

No.

7) Were there any bands apart from the band at 50 kD?

No. A single band.

Read More
Answer

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 having difficulty with this product. Normally our guarantee for 6 months from purchase, but if it was stored properly at -20C in aliquots then we can make an exception as a one time courtesy.

Read More

Question

Hope all is well. Please see customers response below, I have also attached a copy of their answers:

'Thanks for your reply and I have attached my answers for your recommendations.

There are quite a few tests needed to do for this antibody. As specificity for the embryo research, completion of all these suggestion will cost a lot. In fact, our results in ICC have clearly shown the instability of this antibody and our procedures are workable. To catch up with our research progress and save the money, I don’t think we will test so many different procedures. However, your suggestions are very useful for our future research. As all antibodies I bought from Abcam except CREB are at excellent quality I believe this is just an occasional case (may be only for this batch of the antibody). For the anti-CREB antibody, would I ask to get CREDIT for other antibodies if you think my explanations are reasonable? Thanks for your understanding.'

Look forward to your response.

Kind regards,

Read More
Answer

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.

Credit ID: 20123

As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice. Have a good day!

Read More

Question

Thank for the quick response. Please find attached as requested.
Kind regards,

Read More
Answer

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 your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there are only a few tips to provide on this occasion to help improve the results. However, I can recommend you may like to consider trying the following options:

General: 1.

We would always recommend to optimize conditions first on less precious samples.

ICC:

1. We recommend not to mix blocking agents in one experiment as this can affect the results. Also, we would recommend to use 2%, 5% or 10% serum. 30% is quite high. Or you can use 2% or 5% BSA.

2. Could you confirm how long was the fixation in PFA? I suggest to try 10 minutes only if this has not already been tried.

3. I can recommend to fix and then permeabilize. Don’t do both at once.

WB:

1. I can recommend to check the amount of protein loaded for the pre embryo samples.

2. It is unusual that the T47D Cell sample is giving a band at the correct size, but the pre embryo sample is not. I can suggest to review the preparation procedure. Also, I have tried a literature to ensure there is nothing different that would be expected from these sample. I have been unable to find anything, but can suggest you may like to try a more extensive search as this may provide an explanation.

3. I can recommend to try heating to 70oC only when reducing and denaturing as this will help to prevent formation of protein aggregates which could affect the results.

Alternatively, or if these tips do not work, I am pleased to offer you a free of charge replacement or credit note in compensation.

Thank you for your cooperation. I look forward to hearing from you with results from the next experiment and/or details of how you would like to proceed.

Read More

Question

What is the concentration of lot #GR50483-2?

Read More
Answer

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.

Read More

Question

merci de bien vouloir trouver ci-après la liste des anticorps pour commande, je vous serais reconnaissante de bien vouloir m’indiquer pour chaque ref citée le code de remise correspondant afin que je puisse passer la commande ferme.

Par ailleurs, les anticorps référencés ci-dessous, qui ont été acheté à un prix normal, n’ont pas répondu avec le robot SIMON, pourriez-vous envisager un geste commercial pour ces produits qui finalement ont été achetés en vain!

Read More
Answer

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 tester les anticorps listés ci-dessusavec le robot Simon de Protein Simple.Ces codes de réductionsont utilisables sous 120 jours et pour l’achat d’autres anticorps primaires.

Vous devez dans un premier temps soumettre une Abreview avec vos résultats enspécifiant le code promotionnel correspondant à l'anticorps testé. Ce code sera ainsi activé et vous pourrez par la suite contacter notre service clientèle par téléphone pour passer votre prochaine commande, en vous munissant des informations suivantes : code de réduction, numéro de commande de l'anticorps testé.

Concernantles produits pour lesquels vousavez déjà obtenu des résultats, merci dem'envoyer vos données, en échange nous vous attribueronsdes"Abpoints" utilisables pour des remisessur vos commandes ouen bons d'achatAmazon.

Je reste à votre disposition pour plus d’informations concernant cette offre promotionnelle. Dans l’attente de recevoir votre Abreview je vous souhaite unebonne continuation dans vos recherches.

Read More

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