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    products/primary-antibodies/msin3a-antibody-epr27103-53-ab307197.pdf

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Epigenetics and Nuclear Signaling Transcription Other factors
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-mSin3a antibody [EPR27103-53] (ab307197)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-mSin3a antibody [EPR27103-53] (ab307197)
  • Western blot - Anti-mSin3a antibody [EPR27103-53] (ab307197)
  • Western blot - Anti-mSin3a antibody [EPR27103-53] (ab307197)
  • Flow Cytometry (Intracellular) - Anti-mSin3a antibody [EPR27103-53] (ab307197)
  • Flow Cytometry (Intracellular) - Anti-mSin3a antibody [EPR27103-53] (ab307197)
  • Immunoprecipitation - Anti-mSin3a antibody [EPR27103-53] (ab307197)
  • Anti-mSin3a antibody [EPR27103-53] (ab307197)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR27103-53] to mSin3A
  • Suitable for: Flow Cyt (Intra), WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-mSin3a antibody [EPR27103-53]
    See all mSin3A primary antibodies
  • Description

    Rabbit monoclonal [EPR27103-53] to mSin3A
  • Host species

    Rabbit
  • Specificity

    IP: not suitable with mouse.

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IPmore details
    Unsuitable for: ChIP,ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HAP1 whole cell lysate. HeLa, K562, 293T, C6, RAW264.7 and F9 whole cell lysate. Human and mouse testis tissue lysate. Flow Cyt (Intra): Wild-type HAP1 cells. F9 cells. IP: HAP1 whole cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR27103-53
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Co-factors
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of lipids and lipoproteins
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipid metabolism
    • Metabolism
    • Pathways and Processes
    • Cofactors, Vitamins / minerals
    • Co-factors

Associated products

  • Alternative Versions

    • Anti-mSin3a antibody [EPR27103-53] - BSA and Azide free (ab307198)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab307197 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 170 kDa.
IP
1/30.
Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 170 kDa.
IP
1/30.
Application notes
Is unsuitable for ChIP,ICC/IF or IHC-P.

Target

  • Function

    Acts as a transcriptional repressor. Interacts with MXI1 to repress MYC responsive genes and antagonize MYC oncogenic activities. Also interacts with MAD-MAX heterodimers by binding to MAD. The heterodimer then represses transcription by tethering SIN3A to DNA. Acts as a corepressor for REST.
  • Sequence similarities

    Contains 3 PAH (paired amphipathic helix) domains.
  • Post-translational
    modifications

    SUMO1 sumoylated by TOPORS. Probably desumoylated by SENP2.
  • Cellular localization

    Nucleus. Nucleus > nucleolus. Recruited to the nucleolus by SAP30L.
  • Target information above from: UniProt accession Q96ST3 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 25942 Human
    • Entrez Gene: 20466 Mouse
    • Entrez Gene: 363067 Rat
    • Omim: 607776 Human
    • SwissProt: Q96ST3 Human
    • SwissProt: Q60520 Mouse
    • Unigene: 513039 Human
    • Unigene: 15755 Mouse
    • Alternative names

      • AW553200 antibody
      • DKFZP434K2235 antibody
      • FLJ90319 antibody
      • Histone deacetylase complex subunit Sin 3a antibody
      • Histone deacetylase complex subunit Sin3a antibody
      • KIAA0700 antibody
      • Kiaa4126 antibody
      • mKIAA4126 antibody
      • Paired amphipathic helix protein Sin 3a antibody
      • Paired amphipathic helix protein Sin3a antibody
      • Sin 3a antibody
      • SIN3 homolog A antibody
      • SIN3 homolog A transcription regulator (yeast) antibody
      • SIN3 homolog A transcription regulator antibody
      • SIN3 transcription regulator homolog A antibody
      • Sin3a antibody
      • SIN3A protein antibody
      • SIN3A_HUMAN antibody
      • Transcriptional co repressor Sin 3A antibody
      • Transcriptional co repressor Sin3A antibody
      • Transcriptional corepressor Sin3a antibody
      • Transcriptional regulator SIN3A antibody
      see all

    Images

    • Western blot - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      Western blot - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      All lanes : Anti-mSin3a antibody [EPR27103-53] (ab307197) at 1/1000 dilution

      Lane 1 : Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate
      Lane 2 : mSin3A knockout HAP1 whole cell lysate
      Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) at 1/10000 dilution


      Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

      Lysates at 20 µg per lane.

      The samples were run on a Bis-Tris gel.

       

      Performed under reducing conditions.

      False colour image of Western blot: Anti-mSin3A antibody (ab307197) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 (ab8245) loading control staining at 1/20000 dilution, shown in red.

      In Western blot, ab307197 was shown to bind specifically to mSin3A. A band was observed at 170 kDa in wild-type HAP1 cell lysates with no signal observed at this size in mSin3A knockout cell line. To generate this image, wild-type and mSin3A knockout HAP1 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Western blot - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      Western blot - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      All lanes : Anti-mSin3a antibody [EPR27103-53] (ab307197) at 1/1000 dilution

      Lane 1 : Human testis tissue lysate
      Lane 2 : Mouse testis tissue lysate
      Lane 3 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
      Lane 4 : 293T (human embryonic kidney epithelial cell) whole cell lysate
      Lane 5 : F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution


      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      The identity of the bands below 75 kDa are unknown.

      The blot was developed using a higher sensitivity ECL substrate.

      Exposure time: 180 seconds.

    • Western blot - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      Western blot - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      All lanes : Anti-mSin3a antibody [EPR27103-53] (ab307197) at 1/1000 dilution

      Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate 20 µg
      Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution


      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      The identity of the bands below 75 kDa are unknown.

      This blot was developed using a high sensitivity ECL substrate.

      Exposure time: 180 seconds.

    • Flow Cytometry (Intracellular) - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      Flow Cytometry (Intracellular) - Anti-mSin3a antibody [EPR27103-53] (ab307197)

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) (Right panel) compared to mSin3A knockout HAP1 (Left panel) cells labeling mSin3A with AB307197 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

      A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

    • Flow Cytometry (Intracellular) - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      Flow Cytometry (Intracellular) - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labeling mSin3A with AB307197 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
    • Immunoprecipitation - Anti-mSin3a antibody [EPR27103-53] (ab307197)
      Immunoprecipitation - Anti-mSin3a antibody [EPR27103-53] (ab307197)

      mSin3A was immunoprecipitated from 0.35 mg HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate 10 µg with AB307197 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using AB307197 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HAP1 whole cell lysate 10 µg

      Lane 2: AB307197 IP in HAP1 whole cell lysate 10 ug

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab307197 in HAP1 cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 6 seconds.

    • Anti-mSin3a antibody [EPR27103-53] (ab307197)
      Anti-mSin3a antibody [EPR27103-53] (ab307197)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab307197? Please let us know so that we can cite the reference in this datasheet.

    ab307197 has not yet been referenced specifically in any publications.

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