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ChIPAb+ REST
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REFERENCES
Combinatorial patterning of chromatin regulators uncovered by genome-wide location analysis in human cells.
Ram, Oren, et al. (2011) Cell, 147: 1628-39 (2011)
Species Reactivity Key Applications Host Format Antibody Type
H, M, R WB, IP, ChIP, ChIP-seq Rabbit Affinity Purified Polyclonal Antibody
Description:
ChIPAb+ REST - ChIP Validated Antibody and Primer Set
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Trade Name:
Upstate (Millipore)
Product 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.
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Specificity:
Recognizes REST, Mr 120-200 kDa.
Molecular Weight:
120 kDa predicted, 200 kDa observed
Epitope:
a.a. 801-1097
Immunogen:
The REST purified antibody is made against a GST fusion protein corresponding to amino acids 801-1097 of human REST.
Species Reactivity:
Human
Mouse
Rat
Species Reactivity Note:
Human, mouse, rat. Not tested in other species.
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.
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Control:
Includes negative control rabbit IgG antibody and control primers specific for the RE1 site of human Synapsin.
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.
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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, Synaps |