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Merck

17-10456

Sigma-Aldrich

ChIPAb+ REST - ChIP Validated Antibody and Primer Set

from rabbit

Synonym(s):

Chip Antibody and primer set, Neural-restrictive silencer factor ChIP, Neural-restrictive silencer factor, RE1-silencing transcription factor, X2 box repressor, neuron restrictive silencer factor, repressor binding to the X2 box, RE1-silencing transcription factor ChIP, REST ChIP, X2 box repressor ChIP

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32

biological source

rabbit

Quality Level

100

antibody form

purified immunoglobulin

clone

polyclonal

species reactivity

human, rat, mouse

manufacturer/tradename

ChIPAb+
Upstate®

technique(s)

ChIP: suitable
western blot: suitable

NCBI accession no.

NP_005603

UniProt accession no.

Q13127

shipped in

dry ice

General description

120 kDa predicted, 200 kDa observed
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+ REST set includes the REST antibody, a Normal Rabbit IgG, and control primers which amplify a 89 bp region of the human Snap25 promoter. The REST and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of REST associated chromatin.

Immunogen

Epitope: a.a. 801-1097
The REST purified antibody is made against a GST fusion protein corresponding to amino acids 801-1097 of human REST.

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (4e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µg of either Normal Rabbit IgG (Part No. PP64B), or 4 µg Anti-REST (Part No.CS207378) and the Magna ChIP HiSens (Cat. # 17-10460). Successful immunoprecipitation of REST associated DNA fragments was verified by qPCR using ChIP Primers, human Snap25 (Part No. CS207379) as a positive locus, and CDC25 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.

Western Blotting Analysis:
Human HEK293 cell lysate (30 ug) was resolved by electrophoresis, transferred to PVDF membrane and probed with the anti-REST (1:1000 dilution). Proteins were visualized using a secondary antibody conjugated to HRP and a chemiluminescence detection system. Data courtesy of Glen Corson laboratory of Gail Mandel, Oregon Health and Sciences University, Portland, OR.
REST has a predicted molecular weight of 125 kDa. Observed size was ~200 kDa.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology

Histones
This ChIPAb+ REST -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Biochem/physiol Actions

Recognizes REST, MW 125kDa predicted, 200 kDa observed.

Packaging

25 assays per set. Recommended use: ~2 μg antibody per chromatin immunoprecipitation (dependent upon biological context).

Physical form

Anti-REST (Rabbit Polyclonal). One vial containing 100 µL 1.0 mg/mL purified rabbit polyclonal in buffer containing 0.014 M phosphate buffer, pH 7.6, 0.175 M NaCl, 0.07% sodium azide, and 30% glycerol. Store at -20°C.

Normal Rabbit IgG. One vial containing 125 µg Rabbit IgG in 125 µL storage buffer containing 0.05% sodium azide. Store at -20°C.

ChIP Primers, human Snap25. One vial containing 75 μL of 5 μM of each primer specific for human Snap25 promoter. Store at -20°C.
FOR: AGA CTC CTT TGC AGA CAA TTT CCT
REV: CAA CAC AGA AGA TTT CCA CAA GTA GAC
Format: Purified

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 HeLa cells (4e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µg of either Normal Rabbit IgG (Part No. PP64B), or 4 µg Anti-REST (Part No.CS207378) and the Magna ChIP® HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of REST associated DNA fragments was verified by qPCR using ChIP Primers, human Snap25.
Control
Includes normal rabbit IgG and primers specific for human Snap25.

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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Advance your Epigenetics Research

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.

Global Trade Item Number

SKUGTIN
17-1045604053252870392

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