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ChIPAb+ SMRT - ChIP Validated Antibody and Primer Set
Description:
ChIPAb+ SMRT - ChIP Validated Antibody and Primer Set
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+ SMRT set includes the SMRT antibody, a negative control mouse ascites, and qPCR primers which amplify a 299 bp region of human IκBα promoter. The SMRT and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of SMRT-associated chromatin.
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Specificity:
Recognizes SMRT.
Molecular Weight:
~ 275 kDa
Immunogen:
Recombinant protein corresponding to human SMRT.
Isotype:
IgG2aκ
Background Information:
The Silencing mediator of retinoic acid & thyroid hormone receptor protein, commonly called the SMRT protein mediates the transcriptional repression of some nuclear receptors by promoting chromatin condensation, thus preventing access of the basal transcription machinery. Consistent with this activity, this protein is known to form a large corepressor complex containing SIN3A/B and histone deacetylases HDAC1 and HDAC2. SMRT is also a component of the N-Cor repressor (nuclear receptor corepressor), a multi-subunit complex minimally composed of NCOR1, NCOR2, HDAC3, TBL1X, TBL1R, CORO2A and GPS2. SMRT and nuclear receptor corepressor N-CoR are related transcriptional corepressors which contain two distinct domains capable of interacting with unliganded nuclear receptors to repress their basal transcriptional activity.
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Species Reactivity:
Human
Mouse
Rat
Application Notes:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from serum starved, TNFα treated (20 ng/mL, 30 min) HeLa cells (~3 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of Negative Ascites or 2 µL Anti-SMRT) and the Magna ChIP™ G Kit (Cat. # 17-611).
Successful immunoprecipitation of SMRT associated DNA fragments was verified by qPCR using ChIP Primers, IĸBα promoter as a positive locus, and β-Actin promoter primers as a negative locus (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 ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
Human brain lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-SMRT (1:1000 dilution).
Proteins were visualized using a Goat Anti-Mouse IgG conjugated to HRP and a chemiluminescence detection system (Please see figures).
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Control:
Includes negative control mouse ascites and primers specific for human IκBα promoter.
Quality Assurance:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from serum starved, TNFα treated (20 ng/mL, 30 min) HeLa cells (~3 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of Negative Ascites or 2 µL Anti-SMRT) and the Magna ChIP™ G Kit (Cat. # 17-611).
Successful immunoprecipitation |