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Anti-CREB1 PHOSPHO Antibody
Also for CREB1 (NM_004379)
|CREB phospho peptide corresponding to amino acid residues 122-147 of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH).|
|human, mouse, rat
||ELISA: 1:1,000 - 1:6,000, WB: 1:500 - 1:2,000, IHC: 20 ug/ml
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|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.
|Homo sapiens cAMP responsive element binding protein 1 (CREB1), transcript variant A|
EGFR1 Signaling Pathway
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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 (# 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 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.
affinity purified anti-CREB pS133 antibody was used at 20 ?g/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.