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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, M, R | WB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10456 |
| Brand Family | Upstate |
| Trade Name |
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| 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 |
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| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| 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 |
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| 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 |
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| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| 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 |
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| 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 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 |
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| 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). |