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Antibodies

CREB phospho S133 Antibody

Product Sizes
100 ul
£600.00
600-401-270-100UL
About this Product
SKU:
600-401-270
Additional Names:
CREB1|rabbit anti-CREB pS133 Antibody, Cyclic AMP-responsive element-binding protein 1, Active transcription factor CREB antibody, cAMP responsive element binding protein 1 antibody, CREB 1 antibody, CREB-1, CREB1 antibody, MGC9284 antibody, Transactivator protein antibody
Application:
ELISA, Immunohistochemistry, Western Blot
Buffer:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride
CE/IVD:
RUO
Clonality:
Polyclonal
Concentration:
0.064 mg/ml
Extra Details:
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.
Formulation:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Host:
Rabbit
Immunogen:
CREB phospho peptide corresponding to amino acid residues 122-147 of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH).
Isotype:
IgG
Physical State:
Liquid
Reactivities:
Human, Mouse, Rat
Shipping Conditions:
Dry Ice
Specificity:
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
Storage Conditions:
-20[o]C aliquoted. Aliquot. Avoid freeze/thaw cycles., 2-8[o]C diluted. Aliquot. Avoid freeze/thaw cycles.
Supplier:
Rockland Inc
Type:
Antibody: Polyclonal Antibody
Rockland's affinity purified anti-CREB pS133 antibody was used at 20ug/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 at ~46kDa. 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.
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 indicated amounts (100ng, 50ng, 25ng, 12.5ng) of control (-) and in-vitro phosphorylated CREB (P) were loaded to show that the antibody reacts specifically with the phosphorylated form. Blots were blocked in 5% milk in TBS+0.1% Tween-20 (TBST-M) overnight at 4°C. Detection occurs using a 1:500 dilution of antibody diluted in TBST-M and incubated at room temperature with rocking for 1 hour. Blots were rinsed 6X with TBST and incubated with goat anti-rabbit-HRP at 1:5000 in TBST-M at room temperature for 45 min. Blots were again rinsed 6X with TBST and then processed using ECL reagent (Amersham) according to manufacturer's instructions. Exposure time: 1 min using Kodak Biomax MR film. Personal Communication, R. Screaton, The Salk Institute for Biological Studies.