ChIPAb+ Phospho-CREB (Ser133)

ChIPAb+ Phospho-CREB (Ser133)

Stock Code: 3586332
Manufacturer Part No: 17-10131
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Analysis Note


ControlIncludes negative control normal rabbit IgG and primers specific for human cFOS CRE.


Application


Research Sub CategoryTranscription Factors


This ChIPAb+ Phospho-CREB (Ser133) -ChIP Validated Antibody & Primer Set (Antibody ChIP) conveniently includes the antibody & the specific control PCR primers.


Research CategorySignalingEpigenetics & Nuclear Function


Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from 293T cells (5 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 4 µg of Normal Rabbit IgG, (Part No. P64B), or 4 µg of Anti-Phospho-CREB (Ser133) antibody (Part No. CS204400) and the Magna ChIP® A Kit (Cat. # 17-610).
Successful immunoprecipitation of Phospho-CREB (Ser133) antibody associated DNA fragments was verified by qPCR using ChIP Primers cFos CRE (Part No. CS203203) as a positive locus, and cFos downstream 4Kb as a negative locus (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
Untreated (lane 1) and Forskolin-treated (Lane 2) NIH/3T3 lysates were resolved by SDS-PAGE, transferred to PVDF, and probed with anti-phospho-CREB (Ser133) (1:1,000 dilution).
Proteins were visualized using a donkey anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates phosphorylated CREB (Figure 3).
Immunoprecipitation:
10 µL from a representative lot of this antibody was shown to immunoprecipitate phosphorylated CREB from 500 µg of cells treated with 50 mM forskolin.
Immunohistochemistry: A 1:1000 dilution of a representative lot detected phosphorylated CREB in formalin-fixed paraffin-embedded normal and ischemic rat brain sections.
Electrophoretic Mobility Shift:
A representative lot of this antibody has been shown to gel shift by an independent laboratory.


Disclaimer


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.


General description


All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ Phospho-CREB (Ser133) set includes the Phospho-CREB (Ser133) antibody, a negative control normal rabbit IgG, and qPCR primers which amplify a 64 bp region of human cFos CRE

Quality Level100
ManufacturerSIGMA-ALDRICH

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