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Key Specifications Table

Species Reactivity	Key Applications
H, M, R	WB, ChIP

Description
Catalogue Number	17-600
Brand Family	Upstate
Trade Name	ChIPAb+ Upstate
Description	ChIPAb+ CREB - ChIP Validated Antibody and Primer Set, rabbit monoclonal
Overview	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.
Alternate Names	active transcription factor CREB cAMP responsive element binding protein 1 cAMP-response element-binding protein-1 cAMP-responsive element-binding protein 1 transactivator protein
Background Information	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.

Product Information
Components	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
Format	Culture Supernatant
Control	Includes negative control cultured supernatant and primers specific for of human cFos CRE.
Presentation	25 assays per set. Recommended use: ~10 μL antibody per chromatin immunoprecipitation (dependent upon biological context).
Quality Level	MQ100

Applications
Application	This ChIPAb+ CREB -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications	Western Blotting Chromatin Immunoprecipitation (ChIP)
Application Notes	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).

Biological Information
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
Clone	NL904
Host	Rabbit
Specificity	Recognizes CREB, MW ~43 kDa.
Isotype	IgG
Species Reactivity	Human Mouse Rat
Species Reactivity Note	Human, mouse, rat. 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.
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NP_004370
Entrez Gene Summary	This gene encodes a transcription factor that is a memberof 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.
Gene Symbol	CREB1 MGC9284 CREB
UniProt Number	P16220
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 STRUCTURE: 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 LOCATION: Nucleus. PTM: 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. INVOLVEMENT IN DISEASE:A chromosomal aberration involving CREB1 is associated with angiomatoid fibrous histiocytoma (AFH) [MIM:612160]. Translocation t(2;22)(q33;q12) with CREB1 generates a EWSR1/CREB1 fusion gene that is most common genetic abnormality in this tumor type. SEQUENCE SIMILARITIES: Belongs to the bZIP family.Contains 1 bZIP domain.Contains 1 KID (kinase-inducible) domain.
Molecular Weight	~43 kDa

Product Usage Statements

Quality Assurance

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.

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.

Storage and Shipping Information
Storage Conditions	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.

Packaging Information
Material Size	25 assays
Material Package	25 assays per set, ~10 μL per chromatin immunoprecipitation.

Specifications

More Information
SKU	MILL-17-600
Featured	No
Supplier Part Number	17-600
UM	EA
UNSPSC	12352203
Manufacturer Name	Millipore
ProductLine	MILL
Qty	25
MinOrderQty	1
Weight	10.000000
Lead Time	10
Hazardous	N
Energy Star	No
Green	No
Controlled	N

Millipore Chipab+ Creb - Chip Validated Antibody And Primer Set

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