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WB: 20 micrograms of untreated (lane 1) and forskolin treated (lane 2) 3T3 cell lysates were probed with anti-phospho-CREB (1:1000).
Anti-phospho-CREB (Ser133)
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, R, M, Ht | EMSA, IH, IH(P), IP, WB | Rabbit | Purified | Polyclonal Antibody |
Description:
Anti-phospho-CREB (Ser133)
Trade Name:
Upstate (Millipore)
Specificity:
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.
Molecular Weight:
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
Immunogen:
KLH-conjugated synthetic peptide corresponding to the amino acids including and encompassing phosphorylated Ser133 of CREB.
Modifications:
Phosphorylation
Isotype:
IgG
Background Information:
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.
Species Reactivity:
- Human
- Rat
- Mouse
- Hamster
Species Reactivity Note:
Reacts with human, hamster, mouse and rat.
Application Notes:
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.
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.
Control:
Forskolin-stimulated WI-38 nuclear extracts.
Quality Assurance:
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).
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).
Purification Method:
Purified by protein A-Sepharose followed by immunoadsorbant chromatography using an unphosphorylated CREB affinity gel column.
Presentation:
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.
Storage Conditions:
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.
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.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- CREB1
- CREB
- MGC9284
Alternate Names:
- active transcription factor CREB
- cAMP responsive element binding protein 1
- cAMP-response element-binding protein-1
- transactivator protein
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Key Applications:
- Electrophoretic Mobility Shift Assay
- Immunohistochemistry
- Immunohistochemistry (Paraffin)
- Immunoprecipitation
- Western Blotting
Entrez Gene Summary:
This 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, an octameric palindrome. The protein is phosphorylated by several protein kinases, and induces transcription of genes in response to hormonal stimulation of the cAMP pathway. Alternate splicing of this gene results in two transcript variants encoding different isoforms.
UniProt Summary:
FUNCTION:This protein binds the cAMP response element (CRE), a sequence present in many viral and cellular promoters. CREB stimulates transcription on binding to the CRE. Transciption activation is enhanced by the TORC coactivators which act independently of Ser-133 phosphorylation. Implicated in synchronization of circadian rhythmicity.
SUBUNIT: Interacts with PPRC1. Binds DNA as a dimer. This dimer is stabilized by magnesium ions. Interacts, through the bZIP domain, with the coactivators TORC1/CRTC1, TORC2/CRTC2 and TORC3/CRTC3. When phosphorylated on Ser-133, binds CREBBP By similarity.
SUBCELLULAR: Nucleus.
PMT:Stimulated by phosphorylation. Phosphorylation of both Ser-133 and Ser-142 in the SCN regulates the activity of CREB and participates in circadian rhythm generation. Phosphorylation of Ser-133 allows CREBBP binding By similarity. Phosphorylated upon DNA damage, probably by ATM or ATR.
SIMILARITY: Belongs to the bZIP family.
Contains 1 bZIP domain.
Contains 1 KID (kinase-inducible) domain
SUBUNIT: Interacts with PPRC1. Binds DNA as a dimer. This dimer is stabilized by magnesium ions. Interacts, through the bZIP domain, with the coactivators TORC1/CRTC1, TORC2/CRTC2 and TORC3/CRTC3. When phosphorylated on Ser-133, binds CREBBP By similarity.
SUBCELLULAR: Nucleus.
PMT:Stimulated by phosphorylation. Phosphorylation of both Ser-133 and Ser-142 in the SCN regulates the activity of CREB and participates in circadian rhythm generation. Phosphorylation of Ser-133 allows CREBBP binding By similarity. Phosphorylated upon DNA damage, probably by ATM or ATR.
SIMILARITY: Belongs to the bZIP family.
Contains 1 bZIP domain.
Contains 1 KID (kinase-inducible) domain
Brand Family:
Upstate
Product Name:
Anti-phospho-CREB (Ser133)
Concentration:
1 mg/mL
Antibody Type:
Polyclonal Antibody
Qty/Pk:
100 µL
Format:
Purified
Host:
Rabbit