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17-641 ChIPAb+ REST - ChIP Validated Antibody and Primer Set

17-641
25 assays  25 assays per set. Recommended use: ~2 μg antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, M, RWB, IP, ChIP, ChIP-seq
      Description
      Catalogue Number17-641
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ REST - ChIP Validated Antibody and Primer Set
      OverviewAll 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 negative control antibody (purified rabbit IgG), and positive qPCR primers which amplify a 62 bp region near the RE1 site of the human synapsin gene. The REST and negative control antibodies 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
      Background InformationTranscriptional 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. Transcriptional repression by REST-CDYL via the recruitment of histone methyltransferase EHMT2 may be important in transformation suppression.
      References
      Product Information
      FormatAffinity Purified
      Control
      • Includes negative control rabbit IgG antibody and control primers specific for the RE1 site of human Synapsin.
      PresentationAnti-REST (rabbit polyclonal IgG). One vial containing 50 μg of affinity purified antibody in 50 μL PBS with 0.07% sodium azide, and 30% glycerol. Store at -20°C.

      Normal Rabbit IgG. One vial containing 125 μg purified rabbit IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

      ChIP Primers, Synapsin RE1. One vial containing 75 μL of 5 μM of each primer specific for the RE1 site of human Synapsin. Store at -20°C.
      FOR: GGT GCT GAA GCT GGC AGT
      REV: TGG GTT TTA GGA CCA GGA TG
      Quality LevelMQ100
      Applications
      ApplicationREST ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of REST.
      Key Applications
      • Western Blotting
      • Immunoprecipitation
      • Chromatin Immunoprecipitation (ChIP)
      • ChIP-seq
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa S3 cells (4 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 μg of either a normal rabbit IgG or Anti-REST antibody and the EZ-Magna ChIP A Kit (Part No. 17-408). Successful immunoprecipitation of REST-associated DNA fragments was verified by qPCR using primers for CDC25 as a negative control and positive control Primers Syn RE1 (Please see figures). 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:
      HeLa nuclear extract was resolved by electrophoresis, transferred to PVDF membrane and probed with the Ab at a dilution of 1:1000.
      Biological Information
      ImmunogenThe REST purified antibody is made against a GST fusion protein corresponding to amino acids 801-1097 of human REST.
      Epitopea.a. 801-1097
      HostRabbit
      SpecificityRecognizes REST, Mr 120-200 kDa.
      Species Reactivity
      • Human
      • Mouse
      • Rat
      Species Reactivity NoteHuman, mouse, rat. Not tested in other species.
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThis 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
      UniProt SummaryFUNCTION: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 Weight120 kDa predicted, 200 kDa observed
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa S3 cells (4 x 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal rabbit IgG or Anti-REST antibody and the Magna ChIP A Kit (Part No. 17-610) (Please see figures). Successful immunoprecipitation of REST-associated DNA fragments was verified by qPCR using Control Primers.
      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 ConditionsStable 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 Size25 assays
      Material Package25 assays per set. Recommended use: ~2 μg antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      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

      TitleLot Number
      ChIPAb+ REST 2484189
      ChIPAb+ REST - 2383598 2383598
      ChIPAb+ REST - 2445111 2445111
      ChIPAb+ REST - 2460943 2460943
      ChIPAb+ REST - 1951945 1951945
      ChIPAb+ REST - 1992290 1992290
      ChIPAb+ REST - 2040406 2040406
      ChIPAb+ REST - 2142713 2142713
      ChIPAb+ REST - 3202451 3202451
      ChIPAb+ REST - 3228892 3228892

      References

      Reference overviewApplicationPub Med ID
      Specificity protein 1 (Sp1)-dependent activation of the synapsin I gene (SYN1) is modulated by RE1-silencing transcription factor (REST) and 5'-cytosine-phosphoguanine (CpG) methylation.
      Paonessa, F; Latifi, S; Scarongella, H; Cesca, F; Benfenati, F
      The Journal of biological chemistry  288  3227-39  2013

      Show Abstract
      23250796 23250796
      Loss of the repressor REST in uterine fibroids promotes aberrant G protein-coupled receptor 10 expression and activates mammalian target of rapamycin pathway.
      Varghese, BV; Koohestani, F; McWilliams, M; Colvin, A; Gunewardena, S; Kinsey, WH; Nowak, RA; Nothnick, WB; Chennathukuzhi, VM
      Proceedings of the National Academy of Sciences of the United States of America  110  2187-92  2013

      Show Abstract
      Western Blotting23284171 23284171
      Combinatorial patterning of chromatin regulators uncovered by genome-wide location analysis in human cells.
      Ram, Oren, et al.
      Cell, 147: 1628-39 (2011)  2011

      Show Abstract
      Chromatin Immunoprecipitation (ChIP)22196736 22196736

      Brochure

      Title
      Advance your Epigenetics Research

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      Categories

      Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies
      Life Science Research > Antibodies and Assays > Primary Antibodies
      Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies