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17-10456 Sigma-AldrichChIPAb+ REST - ChIP Validated Antibody and Primer Set

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

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Key Applications: Western Blotting, Chromatin Immunoprecipitation (ChIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: Chromatin Biology, Histones

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Key Spec Table

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

Description
Catalogue Number	17-10456
Brand Family	Upstate
Trade Name	ChIPAb+ Upstate
Description	ChIPAb+ REST - ChIP Validated Antibody and Primer Set
Overview	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.
Alternate Names	Neural-restrictive silencer factor RE1-silencing transcription factor X2 box repressor neuron restrictive silencer factor repressor binding to the X2 box

References

Product Information
Format	Purified
HS Code	3002 15 90
Control	Includes normal rabbit IgG and primers specific for human Snap25.
Presentation	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
Quality Level	MQ100

Applications
Application	This ChIPAb+ REST -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: 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.

Biological Information
Immunogen	The REST purified antibody is made against a GST fusion protein corresponding to amino acids 801-1097 of human REST.
Epitope	a.a. 801-1097
Host	Rabbit
Specificity	Recognizes REST, MW 125kDa predicted, 200 kDa observed.
Species Reactivity	Human Mouse Rat
Antibody Type	Polyclonal Antibody
Entrez Gene Number	NP_005603
Entrez Gene Summary	This gene encodes a transcriptional repressor which represses neuronal genes in non-neuronal tissues. It is a member of the Kruppel-type zinc finger transcription factor family. It represses transcription by binding a DNA sequence element called the neuron-restrictive silencer element. The protein is also found in undifferentiated neuronal progenitor cells, and it is thought that this repressor may act as a master negative regular of neurogenesis. Alternatively spliced transcript variants have been described; however, their full length nature has not been determined. [provided by RefSeq].
Gene Symbol	NRSF XBR
UniProt Number	Q13127
UniProt Summary	FUNCTION:Transcriptional repressor which binds neuron-restrictive silencer element (NRSE) and represses neuronal gene transcription in non-neuronal cells. Restricts the expression of neuronal genes by associating with two distinct corepressors, mSin3 and CoREST, which in turn recruit histone deacetylase to the promoters of REST-regulated genes. Mediates repression by recruiting the BHC complex at RE1/NRSE sites which acts by deacetylating and demethylating specific sites on histones, thereby acting as a chromatin modifier. SUBUNIT STRUCTURE: Interacts with SIN3A, SIN3B and RCOR1. SUBCELLULAR LOCATION: NucleusProbable. TISSUE SPECIFICITY: Ubiquitous. Expressed at higher levels in the tissues of the lymphocytic compartment, including spleen, thymus, peripheral blood lymphocytes and ovary. SEQUENCE SIMILARITY:Contains 9 C2H2-type zinc fingers. SEQUENCE CAUTION: The sequence AAA98503.1 differs from that shown. Reason: Frameshift at positions 189, 212 and 1087. The sequence AAH38985.1 differs from that shown. Reason: Miscellaneous discrepancy. Contaminating sequence. Potential poly-A sequence.
Molecular Weight	120 kDa predicted, 200 kDa observed

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Materials Information

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Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	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.
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.

Product Usage Statements

Quality Assurance

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.

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

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Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
17-10456	04053252870392

Documentation

ChIPAb+ REST - ChIP Validated Antibody and Primer Set SDS

Title
Safety Data Sheet (SDS)

ChIPAb+ REST - ChIP Validated Antibody and Primer Set Certificates of Analysis

Title	Lot Number
ChIPAb+ REST - Q2154721	Q2154721
ChIPAb+ REST - 3289363	3289363
ChIPAb+ REST - 3386092	3386092
ChIPAb+ REST - 3474392	3474392
ChIPAb+ REST - 3494163	3494163
ChIPAb+ REST - 3528191	3528191
ChIPAb+ REST - 3823791	3823791
ChIPAb+ REST - 3900895	3900895
ChIPAb+ REST - 4051305	4051305
ChIPAb+ REST -2665779	2665779

Brochure

Title
Advance your Epigenetics Research

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