17-10057 ChIPAb+ SMRT - ChIP Validated Antibody and Primer Set

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17-10057
25 assays  25 assays per set. Recommended use: ~2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, M, R ICC, IHC, IP, WB, EMSA, ChIP
      Description
      Catalogue Number17-10057
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ SMRT - 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+ 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.
      Alternate Names
      • CTG repeat protein 26
      • Silencing mediator of retinoic acid and thyroid hormone receptor
      • T3 receptor-associating factor
      • Thyroid-, retinoic-acid-receptor-associated corepressor
      • nuclear receptor co-repressor 2
      • silencing mediator for retinoid and thyroid hormone receptors
      Background InformationThe 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.
      References
      Product Information
      FormatAscites
      Control
      • Includes negative control mouse ascites and primers specific for human IκBα promoter.
      PresentationAnti-SMRT (mouse monoclonal). One vial containing 50 µL of unpurified mouse monoclonal ascites with 0.05% sodium azide. Store at -20°C.

      Negative Ascites (mouse). One vial containing 50 µL of mouse ascites with 0.05% sodium azide. Store at -20°C.

      ChIP Primers, IκBα promoter. One vial containing 75 μL of 5 μM of each primer specific for human IĸBα promoter. Store at -20°C.
      FOR: GAC GAC CCC AAT TCA AAT CG
      REV: TCA GGC TCG GGG AAT TTC C
      Quality LevelMQ100
      Applications
      ApplicationThe ChIPAb+ SMRT set includes the SMRT antibody, a negative control mouse ascites & qPCR primers which amplify a 299 bp region of human IκBα promoter.
      Key Applications
      • Immunocytochemistry
      • Immunohistochemistry
      • Immunoprecipitation
      • Western Blotting
      • Electrophoretic Mobility Shift Assay
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin 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).
      Biological Information
      ImmunogenRecombinant protein corresponding to human SMRT.
      HostMouse
      SpecificityRecognizes SMRT.
      IsotypeIgG2aκ
      Species Reactivity
      • Human
      • Mouse
      • Rat
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThe protein encoded by this gene interacts with certain retinoid and thyroid hormone receptors involved in transcriptional repression. The encoded protein is part of a large protein complex that facilitates chromatin condensation and repression. Two transcript variants encoding different isoforms have been found for this gene.
      Gene Symbol
      • NCOR2
      • CTG26
      • N-CoR2
      • SMAP270
      • SMRT
      • SMRTE
      • SMRTE-tau
      • TNRC14
      • TRAC
      • TRAC-1
      • TRAC1
      Purification MethodUnpurified
      UniProt Number
      UniProt SummaryFUNCTION: Mediates the transcriptional repression activity of some nuclear receptors by promoting chromatin condensation, thus preventing access of the basal transcription.
      SIZE: 2517 amino acids; 274034 Da
      SUBUNIT: Interacts with HDAC7 (By similarity). Forms a large corepressor complex that contains SIN3A/B and histone deacetylases HDAC1 and HDAC2. This complex associates with the thyroid (TR) and the retinoid acid receptors (RAR) in the absence of ligand, and may stabilize their interaction with TFIIB. The SRMT isoform interacts with HDAC10. Interacts with MINT. Component of the N-Cor repressor complex, at least composed of NCOR1, NCOR2, HDAC3, TBL1X, TBL1R, CORO2A and GPS2. Interacts with CBFA2T3.
      SUBCELLULAR LOCATION: Nucleus.
      TISSUE SPECIFICITY: Ubiquitous. High levels of expression are detected in lung, spleen and brain.
      DOMAIN: SwissProt: Q9Y618 The N-terminal region contains repression functions that are divided into three independent repression domains (RD1, RD2 and RD3). The C-terminal region contains the nuclear receptor- interacting domains that are divided in two separate interaction domains (ID1 and ID2). & The two interaction domains (ID) contain a conserved sequence referred to as the CORNR box. This motif is required and sufficient to permit binding to unligated TR and RARS. Sequences flanking the CORNR box determine nuclear hormone receptor specificity.
      SIMILARITY: Belongs to the N-CoR nuclear receptor corepressors family. & Contains 2 SANT domains.
      Molecular Weight~ 275 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin 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 (Please see figures).

      Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
      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 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+ SMRT - ChIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

      ChIPAb+ SMRT - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ SMRT - 23826512382651
      ChIPAb+ SMRT - 24274982427498
      ChIPAb+ SMRT - 19824931982493
      ChIPAb+ SMRT - 20090162009016
      ChIPAb+ SMRT - 20609872060987
      ChIPAb+ SMRT - 21370472137047
      ChIPAb+ SMRT - NG18274487NG18274487
      ChIPAb+ SMRT - NRG1755667NRG1755667
      ChIPAb+ SMRT -24954992495499
      ChIPAb+ SMRT -26054172605417

      References

      Reference overviewApplicationPub Med ID
      Nuclear corepressors mediate the repression of phospholipase A2 group IIa gene transcription by thyroid hormone.
      Sharma, P; Thakran, S; Deng, X; Elam, MB; Park, EA
      The Journal of biological chemistry  288  16321-33  2013

      Show Abstract
      Western Blotting23629656 23629656
      Knockdown of menin affects pre-mRNA processing and promoter fidelity at the interferon-gamma inducible IRF1 gene.
      Auriemma, LB; Shah, S; Linden, LM; Henriksen, MA
      Epigenetics & chromatin  5  2  2012

      Show Abstract
      22240255 22240255

      Brochure

      Title
      Advance your Epigenetics Research (MilliporeSigma)

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      Categories

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