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Merck
CN

17-600

Sigma-Aldrich

ChIPAb+ CREB - ChIP Validated Antibody and Primer Set, rabbit monoclonal

culture supernatant, clone NL904, from rabbit

Synonym(s):

active transcription factor CREB, cAMP responsive element binding protein 1, cAMP-response element-binding protein-1, cAMP-responsive element-binding protein 1, transactivator protein

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.52

Key Documents

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biological source

rabbit

Quality Level

100

antibody form

culture supernatant

clone

NL904, monoclonal

species reactivity

mouse, rat, human

manufacturer/tradename

ChIPAb+
Upstate®

technique(s)

ChIP: suitable
western blot: suitable

isotype

IgG

NCBI accession no.

NP_004370

UniProt accession no.

P16220

shipped in

dry ice

Gene Information

human ... CREB1(1385)

Related Categories

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+ CREB, antibody/primer set includes the CREB, antibody, a negative control antibody (purified rabbit IgG), and qPCR primers which amplify a 64 bp region flanking the CRE of the human cFos gene. The CREB and negative control cultured supernatant are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of CREB associated chromatin.
Every lot of the ChIPAb+ line of antibodies is individually validated for chromatin precipitation, in order to guarantee successful ChIP assays every time. Each antibody includes a control primer set for performance confirmation. CREB antibody and the negative control antibody (rabbit normal IgG) can be used to demonstrate that the CREB antibody is functionally validated in the precipitation of CREB associated chromatin.
The qPCR primers included flank the CREB binding site in human NR4A2 promoter.
~43 kDa

Immunogen

CREB purified antibody is made against a KLH-conjugated synthetic peptide corre-sponding to amino acids 5-24 of human CREB.
Epitope: a.a. 5-24

Application

Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from 293T cells (5 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 10 µL of either a negative supernatant, or CREB antibody and the Magna ChIP® A Kit (Cat. # 17-610).
Successful immunoprecipitation of CREB-associated DNA fragments was verified by qPCR using cFos downstream 4Kb (negative) and cFos CRE (positive) (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 A ChIP (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Representative lot data.
HeLa nuclear extract was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-CREB, clone NL904 (1:8,000 dilution)
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates CREB (~43 kDa).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
This ChIPAb+ CREB -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Biochem/physiol Actions

Not tested in other species. The immunogen sequence is identical in a wide range of animal and plant species, so broad cross-reactivity is expected.
Recognizes CREB, MW ~43 kDa.

Packaging

25 assays per set, ~10 μL per chromatin immunoprecipitation.

Physical form

25 assays per set. Recommended use: ~10 μL antibody per chromatin immunoprecipitation (dependent upon biological context).

Preparation Note

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, 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.

Analysis Note

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from 293T cells (5 X 106 cell equivalents per IP) were subjected to chromatin immuno-precipitation using 10 µL of either a negative control cultured supernatant, or CREB antibody and the Magna ChIP® A Kit (Cat. # 17-610). Successful immunoprecipitation of CREB-associated DNA fragments was verified by qPCR using Control Primers (Figure 1).
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Control
Includes negative control cultured supernatant and primers specific for of human cFos CRE.

Other Notes

Anti-CREB (Rabbit monoclonal IgG): Part No. CS203204. One vial containing 250 μL of cultured supernatant containing 0.05% sodium azide. Store at -20°C.
Negative Control Supernatant: Part No. CS203202. One vial containing 250 μL of cultured supernatant containing 0.05% sodium azide. Store at -20°C.
ChIP Primers cFos CRE: Part No. CS203203. One vial containing 75 μL of 5 μM of each primer specific for the cFos CRE. Store at -20°C.
FOR: GGC CCA CGA GAC CTC TGA GAC A
REV: GCC TTG GCG CGT GTC CTA ATC T

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

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.

Storage Class Code

10 - Combustible liquids

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Soren Impey et al.
Cell, 119(7), 1041-1054 (2004-12-29)
The CREB transcription factor regulates differentiation, survival, and synaptic plasticity. The complement of CREB targets responsible for these responses has not been identified, however. We developed a novel approach to identify CREB targets, termed serial analysis of chromatin occupancy (SACO)
Chunpeng Zhang et al.
Cancer research, 71(21), 6738-6748 (2011-09-09)
The enhancer pioneer transcription factor FoxA1 is a global mediator of steroid receptor (SR) action in hormone-dependent cancers. In castration-resistant prostate cancer (CRPC), FoxA1 acts as an androgen receptor cofactor to drive G₂ to M-phase cell-cycle transit. Here, we describe
Alejandro Maureira et al.
Journal of cellular biochemistry, 117(8), 1797-1805 (2016-01-06)
Proper regulation of gene expression is essential for normal development, cellular growth, and differentiation. Differential expression profiles of mRNA coding for vertebrate Ric-8B during embryo and adult stages have been observed. In addition, Ric-8B is expressed in few cerebral nuclei
Kristen M Meldi et al.
Endocrinology, 153(10), 4905-4917 (2012-08-07)
The transition from follicle to corpus luteum after ovulation is associated with profound morphological and functional changes and is accompanied by corresponding changes in gene expression. The gene encoding the α subunit of the dimeric reproductive hormone inhibin is maximally
J W Dalley et al.
The European journal of neuroscience, 11(4), 1265-1274 (1999-04-02)
The present study investigated the effects of excitotoxic lesions of the prefrontal cortex (PFC) on dopamine (DA) and excitatory amino acid (EAA) function in the nucleus accumbens core using in vivo microdialysis in freely moving rats. As a postsynaptic marker
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Epigenetics describes heritable changes in gene expression caused by non-genetic mechanisms. Epigenetic regulation allows a cell to vary its response based on its biological and environmental contexts. Epigenetic changes can effect transcriptional and post-transcriptional regulation via mechanisms such as histone modification, chromatin and nucleosome remodeling, DNA methylation, and small and non-coding RNA-mediated regulation. These mechanisms, in cooperation with transcription factors and other nucleic acid-binding proteins, regulate gene expression. Epigenetic mechanisms of gene regulation impacts diverse areas of research—from agriculture to human health. Common epigenetic assays such as chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) rely on high quality antibodies that recognize specific epigenetic modifications for accurate results. EMD Millipore offers over 100 ChIPAb+™ and RIPAb+™ validated antibody kits that are quality tested on ChIP/RIP assays and are conveniently provided with control qPCR primers and negative control antibodies to ensure first time ChIP/RIP success.

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