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Primary Antibodies  >  Transcription Factor Antibodies
 

CREB phospho S133 Antibody

Rabbit Polyclonal
Code

600-401-270
Size

100 µL
Price

$309.00
Availability

Ships next business day.
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E-mail: orders@rockland-inc.com
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Rockland's affinity purified anti-CREB Ps133 antibody was used at 20 µg/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows moderate to strong nuclear staining of tonsillar lymphocytes.  Tissue was formalin-fixed and paraffin embedded.  The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.  Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.
Anti-CREB pS133 was used to detect phosphorylated CREB by western blot.  Recombinant His-tagged human CREB was produced in E.coli and purified by metal affinity chromatography.  An aliquot of purified CREB was phosphorylated in-vitro using Protein Kinase A and ATP.  western blot of control (-) and in-vitro phosphorylated CREB (+) was used to show that the antibody reacts specifically with the phosphorylated form.  Pan reactive CREB (Rockland # 100-401-195) reacts equally with both non-phosphorylated and phosphorylated CREB (not shown).  Detection occurs using a 1:500 dilution of antibody followed by 1:5,000 dilution of HRP Goat-a-Rabbit IgG with visualization via ECL.  Film exposure was approximately 1’.  Other detection systems will yield similar results. Personal Communication, Boss, J., Emory University School of Medicine, Atlanta, GA.
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Synonyms: Active transcription factor CREB antibody, cAMP response element binding protein antibody, cAMP responsive element binding protein 1 antibody, CREB 1 antibody, CREB1 antibody, MGC9284 antibody, Transactivator protein antibody

CREB phospho S133 Antibody Properties

Description
Anti-CREB pS133 (RABBIT) Antibody - 600-401-270
Host
Rabbit
Target Species
Human
Known Cross Reactivity
human, mouse, rat
Clonality
Polyclonal
Application
ELISA : 1:1,000 - 1:6,000
Western Blot : 1:500 - 1:2,000
Immunochemistry: 20 µg/ml
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG
Label
Concentration
0.064 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
CREB1
Download CofA
Lot#  

CREB phospho S133 Antibody Description

Background
The CREB (Cyclic AMP-response-element-binding-protein 1) gene encodes a transcription factor that is a member of the leucine zipper family of DNA binding proteins. This protein binds as a homodimer to the cAMP-responsive element (CRE element TGANNTCA), an octameric palindrome. Phosphorylation by cAMP-dependent protein kinase (PKA) at serine-119 is required for interaction with DNA and phosphorylation at serine-133 allows CREB to interact with CBP (CREB binding protein) leading to interaction with RNA polymerase II.  Alternate splicing of this gene results in two transcript variants encoding different isoforms.
Immunogen
CREB phospho peptide corresponding to amino acid residues 122-147 of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH).
Immunogen Type
Peptide
Post Translational Modification
Phosphorylation
Storage Condition
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
Anti-CREB pS133 antibody reacts with phosphorylated human CREB at pS133 and shows minimal reactivity by western blot with non-phosphorylated CREB and minimal reactivity (1%) by ELISA against the non-phosphorylated form of the immunizing peptide.  This antibody was assayed against a variety of tissues including fibroblasts and B-cell lysates.  Bands of 46 and 43 kDa corresponding to phosphorylated CREB are observed in western blots.  Anti-CREB pS133 is suitable for the detection by immunoblot of phosphoryl-ated human, mouse and rat CREB.  No cross-reactivity occurs with non-phosphorylated CREB.  For immunohistochemistry, formalin fixed, paraffin embedded human tissue shows moderate to strong nuclear staining in a variety of cells with minimal background staining.  Although not tested this antibody is likely functional by FACS and immunoprecipitation.
Purity/Specifity
Anti-CREB pS133 was affinity purified from monospecific antiserum by immunoaffinity purification.  Antiserum was first sequentially pre-adsorbed against a control E.coli lysate, nickel-purified recombinant CREB and the non-phosphorylated form of the immunizing peptide.  The resultant depleted antiserum was then purified against the phosphorylated form of the immunizing peptide.  This phospho specific polyclonal antibody is specific for phosphorylated pS133 of human CREB.  Reactivity with non-phosphorylated CREB is minimal. This antibody does show cross reactivity with pS133 phosphorylated CREB from mouse and rat.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Moreno, C.S., Beresford, G., Louis-Plence, P., Morris, A.C., and Boss, J.M. (1999) CREB regulates MHC class II expression in a CIITA dependent manner. Immunity 10: 143-151. Arthur JS and Cohen, P. (2000) MSK1 is required for CREB phosphorylation in response to mitogens in mouse embryonic stem cells. FEBS Lett, 482(1-2): 44-8. Kwok RP, Laurance ME, Lundblad JR, Goldman PS, Shih H, Connor LM, Marriott SJ, Goodman RH. (1996) Control of cAMP-regulated enhancers by the viral transactivator Tax through CREB and the co-activator CBP. Nature 380:642-646.
Specific Reference
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Product Type
Primary Antibodies;
Applications
ELISA, WB, GS, Other
Reacts With
human, mouse, rat
Catalog Number
100-401-195

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human
Catalog Number
100-401-218

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF, FC
Reacts With
human, mouse, rat, chicken
Catalog Number
100-401-401

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human, mouse, rat, dog, orangutan
Catalog Number
600-401-873

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry