Western Blotting (WB), Immunoprecipitation (IP), Enzyme Immunoassay (EIA)
Sequence
Q-K-R-R-E-I-L-S-R-R-P-pS-Y-R-K-I-L-N-D-L-S-S-D-A-P correspond ing to aa 122-147
Specificity
This phospho specific polyclonal antibody reacts with phosphorylated human CREB at pS133 and shows minimal reactivity by western blot with non-phosphorylated CREB and minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide. This product 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.
Purification
Affinity Purified
Immunogen
CREB phospho peptide corresponding to a region of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH).
Western Blot: 1: 1,000. ELISA. Immunoprecipitation. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Restrictions
For Research Use only
Format
Liquid
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 containing 0.1 % (w/v) Sodium Azide
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing.
Storage
4 °C/-20 °C
Storage Comment
Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
Target
CREB1
(cAMP Responsive Element Binding Protein 1 (CREB1))
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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 cAMPdependent 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.Synonyms: CREB-1, Cyclic AMP-responsive element-binding protein 1, cAMP-responsive element-binding protein 1