For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

We use cookies to make our site as useful as possible.

Our Cookie Policy explains how you can opt-out of the cookies we use.

If you continue without changing your cookie settings, we'll assume you’re happy with this.

Continue Continue

United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Diagnostic & Therapeutic Solutions
    Custom solutions & partnerships

    Custom antibody development and commercial partnerships to advance your diagnostic and therapeutic discovery.

    Create custom solutions with us

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

Supporting our customers and employees during the COVID-19 pandemic. Read more

  1. Link

    creb-phospho-s133-antibody-e113-ab32096.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Epigenetics and Nuclear Signaling Transcription Domain Families HLH / Leucine Zipper HLH / Leucine Zipper
Share by email
RecombinantRabMAb

Recombinant Anti-CREB (phospho S133) antibody [E113] (ab32096)

  • Datasheet
  • SDS
Reviews (13)Q&A (8)References (95)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Flow Cytometry - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] (ab32096)
  • Anti-CREB (phospho S133) antibody [E113] (ab32096)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E113] to CREB (phospho S133)
  • Suitable for: WB, IHC-P, IP, Flow Cyt, ICC/IF
  • Reacts with: Mouse, Rat, Human

You may also be interested in

IHC
Normal Horse Serum (sterile) (ab139501)
Biochemical
H-89, PKA inhibitor (ab143787)
ELISA
Product image
CREB1 pS133 + CREB1 ELISA Kit (ab176659)

View more associated products

Overview

  • Product name

    Anti-CREB (phospho S133) antibody [E113]
    See all CREB primary antibodies
  • Description

    Rabbit monoclonal [E113] to CREB (phospho S133)
  • Host species

    Rabbit
  • Specificity

    This antibody is specific for CREB phosphorylated on Serine 133. The immunogen of the antibody shares 94%homology with CREB (S136) and 86% homology with ATF1 (pS63). No experiment has been performed to verify the possible cross-reactivity.
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human CREB aa 100-200. The exact sequence is proprietary.
    Database link: P16220
    (Peptide available as ab182754)

  • Positive control

    • WB: A431 cell lysate; HeLa treated with anisomycin for 30 minutes ICC/IF: A431 and HeLa IHC-P: Thyroid gland adenocarcinoma. IP: HeLa treated with 25 ug/mL anisomycin for 30 minutes.
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E113
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • HLH / Leucine Zipper
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Immunology
    • Innate Immunity
    • TLR Signaling

Associated products

  • Alternative Versions

    • HRP Anti-CREB antibody [E113] (ab219741)
    • Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (ab220798)
  • Assay kits

    • Total CREB Transcription Factor Assay Kit (Colorimetric) (ab207201)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Rat CREB protein (ab43602)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • (S)-3,5-DHPG, group I mGlu agonist (ab120007)
    • (R,S)-CHPG, mGlu5 agonist (ab120039)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32096 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Mouse
Rat
Human
All applications
Chicken
Cow
Zebrafish
Application Abreviews Notes
WB (7)
1/5000. Predicted molecular weight: 37 kDa.

For unpurified use at 1/500.

IHC-P (3)
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

IP
1/40 - 1/100.

 

Flow Cyt
Use at an assay dependent concentration.
ICC/IF (1)
1/100 - 1/250.
Notes
WB
1/5000. Predicted molecular weight: 37 kDa.

For unpurified use at 1/500.

IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

IP
1/40 - 1/100.

 

Flow Cyt
Use at an assay dependent concentration.
ICC/IF
1/100 - 1/250.

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.
  • Target information above from: UniProt accession P16220 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 281713 Cow
    • Entrez Gene: 1385 Human
    • Entrez Gene: 12912 Mouse
    • Entrez Gene: 81646 Rat
    • Omim: 123810 Human
    • SwissProt: P27925 Cow
    • SwissProt: P16220 Human
    • SwissProt: Q01147 Mouse
    • SwissProt: P15337 Rat
    • Unigene: 516646 Human
    • Unigene: 453295 Mouse
    • Unigene: 90061 Rat
    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 (phospho S133) antibody [E113] (ab32096)
    Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    All lanes : Anti-CREB (phospho S133) antibody [E113] (ab32096) at 1/500 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates
    Lane 2 : HeLa whole cell lysates treated with anisomycin at 25ug/ml for 30 minutes
    Lane 3 : HeLa whole cell lysates treated with anisomycin at 25ug/ml for 30 minutes, then the membrane was incubated with phosphatase

    Lysates/proteins at 15 µg per lane.

    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 and concentration: 5% NFDM/TBST

  • Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] (ab32096)

    Immunocytochemistry/Immunofluorescence analysis of LP treated and untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) labelling CREB with purified ab32096 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain at a dilution of 1/200. 

    Confocal image showing nuclear staining on HeLa cells .The signal decreased after Lambda Protein Phosphatase treatment ( 31℃,2h).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue labelling CREB with purified ab32096 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
  • Flow Cytometry - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Flow Cytometry - Anti-CREB (phospho S133) antibody [E113] (ab32096)

    Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling CREB with purified ab32096 at 1/70 dilution(10µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] (ab32096)

    ab32096 at 1/100 dilution immunoprecipitating CREB (phospho S133) in HeLa (human cervix adenocarcinoma) treated with 25ug/mL anisomycin for 30 minutes, whole cell lysate, observed at 40 kDa (lanes 1 and 2).

    Lane 1 (input): HeLa treated with 25ug/mL anisomycin for 30 minutes. Whole cell lysate, 10μg.

    Lane 2 (+): ab32096 + HeLa treated with 25ug/mL anisomycin for 30 minutes. Whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32096 in treated with 25ug/mL anisomycin for 30 minutes. Whole cell lysate

    For western blotting, ab32096 at 1/200 dilution followed by ab131366 VeriBlot for IP Detection Reagent (HRP) at 1/1000 for detection.

    Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    All lanes : Anti-CREB (phospho S133) antibody [E113] (ab32096) at 1/200 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 250ng/ml anisomycin for 30 minutes whole cell lysates
    Lane 2 : Mouse brain lysates
    Lane 3 : Rat brain lysates
    Lane 4 : Rat cerebellum lysate
    Lane 5 : Mouse hippocampus lysate
    Lane 6 : Rat hippocampus lysate
    Lane 7 : Rat cerebral cortex lysate

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 37 kDa


    Exposure time: 3 seconds


    Blocking and diluting buffer: 5% NFDM/TBST

    This antibody can`t detect signal in mouse and rat brain related tissues.

  • Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] (ab32096)

    Immunocytochemistry/Immunofluorescence analysis of A431(human epidermoid carcinoma) cells +/- AP 37℃ 1h labelling CREB with purified ab32096 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] (ab32096)This image is courtesy of an Abreivew submitted by Akiko Shingo.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat hippocampus tissue labelling CREB with unpurified ab32096 at 1/100 dilution. Sections were subjected to antigen retrieval by autoclave prior to blocking with 8% milk for 30 minutes at 37°C. The primary antibody was diluted 1/100 with DAKO antibody diluent and incubated with the sample for 18 hours at 4°C. An LSAB-labeled Streptavidin-Biotin conjugated Goat polyclonal antibody was used undiluted as the secondary antibody.

    See Abreview

  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)

    SK-N-SH cells were incubated at 37&degC for 30 minutes with vehicle control (0 &microM) and different concentrations of (S)-3,5-DHPG (ab120007). Increased expression of CREB (phospho S133) in SK-N-SH cells correlates with an increase in (S)-3,5-DHPGconcentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20&microg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with unpurified ab32096 at 1/500 dilution and ab32515 at 1 &microg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.

     

  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Lane 1 : Anti-CREB (phospho S133) antibody [E113] (ab32096) at 1/5000 dilution (purified)
    Lane 2 : purified at 1/5000 dilution

    Lane 1 : Untreated C6 cell lysate
    Lane 2 : C6 treated with Calyculin A cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 37 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    All lanes : Anti-CREB (phospho S133) antibody [E113] (ab32096) at 1/5000 dilution (purified)

    Lane 1 : Untreated NIH/3T3 cell lysate
    Lane 2 : NIH/3T3 treated with Calyculin A

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 37 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    All lanes : Anti-CREB (phospho S133) antibody [E113] (ab32096) at 1/5000 dilution (purified)

    Lane 1 : Untreated HeLa cell lysate
    Lane 2 : HeLa treated with Calyculin A cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 37 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Western blot - Anti-CREB (phospho S133) antibody [E113] (ab32096)

    SK-N-SH cells were incubated at 37&degC for 30 minutes with vehicle control (0 &microM) and different concentrations of (R,S)-CHPG (ab120039). Increased expression of CREB (phospho S133) in SK-N-SH cells correlates with an increase in (R,S)-CHPG concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20&microg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with unpurified ab32096 at 1/500 dilution and ab32515 at 1 &microg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051 ) at 1/10000 dilution and visualised using ECL development solution.

  • Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] (ab32096)

    ab32096 (purified) at 1/50 immunoprecipitating CREB (phospho S133) in HeLa whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/1,500) was used for detection. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] (ab32096)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland adenocarcinoma tissue labelling CREB with unpurified ab32096 at 1/250 dilution.

    Panel A: Cells are untreated. 
    Panel B: Cells are treated with Phosphatase.

  • Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] (ab32096)

    Immunocytochemistry/Immunofluorescence analysis of A431 cells labelling CREB with unpurified ab32096 at 1/250.

    Panel A: Cells are untreated. 

    Panel B: Cells are treated with Phosphatase.

  • Anti-CREB (phospho S133) antibody [E113] (ab32096)
    Anti-CREB (phospho S133) antibody [E113] (ab32096)

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

Datasheets and documents

    • Datasheet
    • SDS
  • References (95)

    Publishing research using ab32096? Please let us know so that we can cite the reference in this datasheet.

    ab32096 has been referenced in 95 publications.

    • Zhong Y  et al. Berberine Attenuates Hyperglycemia by Inhibiting the Hepatic Glucagon Pathway in Diabetic Mice. Oxid Med Cell Longev 2020:6210526 (2020). PubMed: 31976031
    • Pech-Pool S  et al. Thyrotropin-Releasing Hormone (TRH) and Somatostatin (SST), but not Growth Hormone-Releasing Hormone (GHRH) nor Ghrelin (GHRL), Regulate Expression and Release of Immune Growth Hormone (GH) from Chicken Bursal B-Lymphocyte Cultures. Int J Mol Sci 21:N/A (2020). PubMed: 32093298
    • Wu Y  et al. Intensity-dependent effects of consecutive treadmill exercise on spatial learning and memory through the p-CREB/BDNF/NMDAR signaling in hippocampus. Behav Brain Res 386:112599 (2020). PubMed: 32184158
    • Pan B & Deng C Modulation by chronic antipsychotic administration of PKA- and GSK3ß-mediated pathways and the NMDA receptor in rat ventral midbrain. Psychopharmacology (Berl) N/A:N/A (2019). PubMed: 31053935
    • Gu Z  et al. MiR-124 suppression in the prefrontal cortex reduces depression-like behavior in mice. Biosci Rep 39:N/A (2019). PubMed: 31431514
    View all Publications for this product

    Customer reviews and Q&As

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

    1-10 of 21 Abreviews or Q&A

    Western blot abreview for Anti-CREB (phospho S133) antibody [E113]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (mouse pancreatic cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    mouse pancreatic cells
    Blocking step
    odyssey blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 01 2017

    Western blot abreview for Anti-CREB (phospho S133) antibody [E113]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hela)
    Gel Running Conditions
    Reduced Denaturing
    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

    Dr. Lin Zhu

    Verified customer

    Submitted Sep 04 2017

    Western blot abreview for Anti-CREB (phospho S133) antibody [E113]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (BRAIN)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    BRAIN
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
    Read More

    Kyungjoo Seong

    Verified customer

    Submitted Jun 20 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CREB (phospho S133) antibody [E113]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Adult brain including cerebellum)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH 6
    Permeabilization
    No
    Specification
    Adult brain including cerebellum
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Estela Munoz

    Verified customer

    Submitted Dec 17 2015

    Western blot abreview for Anti-CREB (phospho S133) antibody [E113]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Adult Cerebellum)
    Gel Running Conditions
    Reduced Denaturing (12.5% SDS-PAGE Gel)
    Loading amount
    30 µg
    Specification
    Adult Cerebellum
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Read More

    Dr. Estela Munoz

    Verified customer

    Submitted Dec 17 2015

    Immunohistochemistry (Frozen sections) abreview for Anti-CREB (phospho S133) antibody [E113]

    Good
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Permeabilization
    Yes - TritonX-100
    Specification
    Brain
    Blocking step
    Donkey Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 02 2015

    Western blot abreview for Anti-CREB (phospho S133) antibody [E113]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Mouse Neuroblastoma (Neuro2A))
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    400000 cells
    Treatment
    10% FBS and fibroblast growth factor
    Specification
    Mouse Neuroblastoma (Neuro2A)
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 27 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CREB (phospho S133) antibody [E113]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 29°C
    Sample
    Mouse Cell (GL261-mouse glioma)
    Specification
    GL261-mouse glioma
    Permeabilization
    Yes - 0.3% TritonX-100 in 1X TBS
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 28 2014

    Western blot abreview for Anti-CREB (phospho S133) antibody [E113]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Non-reduced Denaturing
    Sample
    Cow Tissue lysate - whole (Subcutaneous Adipose)
    Specification
    Subcutaneous Adipose
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 08 2013

    Western blot abreview for Anti-CREB (phospho S133) antibody [E113]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (8)
    Sample
    Rat Cell lysate - whole cell (Primary hippocampal neurons)
    Specification
    Primary hippocampal neurons
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 14 2013

    1-10 of 21 Abreviews or Q&A

    •  Previous
    • 1
    • 2
    • 3
    • Next 

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Get resources and offers direct to your inbox Sign up
    A-Z by research area
    • Cancer
    • Cardiovascular
    • Cell biology
    • Developmental biology
    • Epigenetics & Nuclear signaling
    • Immunology
    • Metabolism
    • Microbiology
    • Neuroscience
    • Signal transduction
    • Stem cells
    A-Z by product type
    • Primary antibodies
    • Secondary antibodies
    • Biochemicals
    • Isotype controls
    • Flow cytometry multi-color selector
    • Kits
    • Loading controls
    • Lysates
    • Peptides
    • Proteins
    • Slides
    • Tags and cell markers
    • Tools & Reagents
    Help & support
    • Support
    • Make an Inquiry
    • Protocols & troubleshooting
    • Placing an order
    • RabMAb products
    • Biochemical product FAQs
    • Training
    • Browse by Target
    Company
    • Corporate site
    • Investor relations
    • Company news
    • Careers
    • About us
    • Blog
    Events
    • Tradeshows
    • Conferences
    International websites
    • abcam.cn
    • abcam.co.jp

    Join with us

    • LinkedIn
    • facebook
    • Twitter
    • YouTube
    • Terms of sale
    • Website terms of use
    • Cookie policy
    • Privacy policy
    • Legal
    • Modern slavery statement
    © 1998-2021 Abcam plc. All rights reserved.