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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, M, R | WB, IP, ChIP, ChIP-seq |
| Description | |
|---|---|
| Catalogue Number | 17-641 |
| Brand Family | Upstate |
| Trade Name |
|
| 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 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 |
|
| Background Information | 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. Transcriptional repression by REST-CDYL via the recruitment of histone methyltransferase EHMT2 may be important in transformation suppression. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
|
| Presentation | Anti-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 Level | MQ100 |
| Applications | |
|---|---|
| Application | REST ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of REST. |
| Key Applications |
|
| Application Notes | Chromatin 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 | |
|---|---|
| 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, Mr 120-200 kDa. |
| Species Reactivity |
|
| Species Reactivity Note | Human, mouse, rat. Not tested in other species. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
|
| 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 |
|
| UniProt Number |
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| 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 |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin 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 |
|
| 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). |