Anti-phospho-CREB (Ser133) Antibody

CREB is a β-ZIP transcription factor that activates target genes through cAMP response elements. CREB is able to mediate signals from numerous physiological stimuli, resulting in regulation of a broad array of cellular responses. While CREB is expressed in numerous tissues, it plays a large regulatory role in the nervous system. CREB is believed to play a key role in promoting neuronal survival, precursor proliferation, neurite outgrowth and neuronal differentiation in certain neuronal populations. CREB promotes outgrowth and differentiation as a mediator of neurotrophin pathways. CREB is able to selectively activate numerous downstream genes through interactions with different dimerization partners. CREB is activated by phosphorylation at Ser133 by various signaling pathways including Erk, Ca2+ and stress signaling.

Detects CREB only when phosphorylated on Ser133 (~43 kDa). May recognize ATF-1 (~38 kDa) and CREM (~30 kDa) as the two proteins share significant homology to the phosphorylated immunizing peptide.

Epitope: phospho-Ser133

KLH-conjugated synthetic peptide corresponding to the amino acids including and encompassing phosphorylated Ser133 of CREB.

Immunoprecipitation:
10 μL of a previous lot immunoprecipitated phosphorylated CREB from 500 μg of cells treated with 50 μM forskolin.

Immunohistochemistry:
A 1:1000 dilution of previous lots detected phosphorylated CREB in formalin-fixed paraffin-embedded normal and ischemic rat brain sections.

Gel Shift (EMSA):
This antibody has been shown to gel shift by independent labs.

Research Category
Signaling

Research Sub Category
Nuclear Receptors

Use Anti-phospho-CREB (Ser133) Antibody (rabbit polyclonal antibody) is highly published and validated in EMSA, IHC,IHC(P), IP, WB to detect phospho-CREB (Ser133) also known as active transcription factor CREB, cAMP responsive element binding protein 1.

Recognizes p43 phosphorylated CREB. May also recognize p30 and p38 kDa proteins. The p30 & p38 proteins may be phosphorylated CREM and ATF-1 respectively, since the two proteins share significant homology to the phosphorylated CREB immunizing peptide.

Format: Purified

Purified by protein A-Sepharose followed by immunoadsorbant chromatography using an unphosphorylated CREB affinity gel column.

Purified rabbit IgG in buffer containing 0.1 M Tris-Glycine (pH 7.4) 15 mM NaCl, and 0.05% sodium azide with 30% glycerol.

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C.
Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Control
Forskolin-stimulated WI-38 nuclear extracts.

Evaluated by western blot on lysates from mouse NIH/3T3 cells treated with 50 mM forskolin.

Western Blot Analysis: This antibody detected CREB when phosphorylated on Ser133 at 1:1,000 dilution in forskolin treated NIH/3T3 lysates via SDS-PAGE and transferred to PVDF (Immobilon-P).

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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