Overview

  • Product nameAnti-BAF53A antibody - ChIP Grade
    See all BAF53A primary antibodies
  • Description
    Rabbit polyclonal to BAF53A - ChIP Grade
  • SpecificityIn Western blot, ab3882 detects BAF53 and Tip60-TAP in total extracts of Tip60-TAP. Following TEV elution of Tip60-TAP ab3882 only detects BAF53.
  • Tested applicationsSuitable for: ChIP, ICC/IF, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human BAF53A.

    (Peptide available as ab13770.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: NIH 3T3 (Mouse embryonic fibroblast cell line) MEF1 (Mouse embryonic fibroblast cell line) PC12 (Rat adrenal pheochromocytoma cell line) This antibody gave a positive signal in the following tissue lysates: Skeletal Muscle (Mouse) Ovary (Mouse) - normal tissue

Properties

Applications

Our Abpromise guarantee covers the use of ab3882 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
ChIP
ICC/IF
WB
IHC-Fr
  • Application notesChIP: Use 8µl for 2x106 cells.
    ICC/IF: Use at an assay dependent concentration.
    IHC-Fr: 1/400.
    WB: 1µg/ml. Detects a band of approximately 53 kDa. Can be blocked with BAF53A peptide (ab13770).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • RelevanceThis protein is a family member of actin-related proteins (ARPs), which share significant amino acid sequence identity to conventional actins. Both actins and ARPs have an actin fold, which is an ATP-binding cleft, as a common feature. The ARPs are involved in diverse cellular processes, including vesicular transport, spindle orientation, nuclear migration and chromatin remodeling. This protein is a 53 kDa subunit of the BAF (BRG1/brm-associated factor) complex in mammals, which is functionally related to SWI/SNF complex in S. cerevisiae and Drosophila; the latter is thought to facilitate transcriptional activation of specific genes by antagonizing chromatin-mediated transcriptional repression. Together with beta-actin, it is required for maximal ATPase activity of BRG1, and for the association of the BAF complex with chromatin/matrix. It is required for maximal ATPase activity of SMARCA4/BRG1 and for association of the SMARCA4/BRG1 containing remodelling complex BAF with chromatin/nuclear matrix. It is a component of the NuA4 histone acetyltransferase (HAT) complex which is involved in transcriptional activation of select genes principally by acetylation of nucleosomal histone H4 and H2A. This modification may both alter nucleosome - DNA interactions and promote interaction of the modified histones with other proteins which positively regulate transcription. This complex may be required for the activation of transcriptional programs associated with oncogene and proto-oncogene mediated growth induction, tumor suppressor mediated growth arrest and replicative senescence, apoptosis, and DNA repair. NuA4 may also play a direct role in DNA repair when recruited to sites of DNA damage.
    • Cellular localizationNuclear
    • Database links
    • Alternative names
      • 53 kDa BRG1 associated factor A antibody
      • Actin like 6A antibody
      • Actin like protein 6A antibody
      • Actin related protein antibody
      • Actin related protein Baf53a antibody
      • actin-related protein 4 antibody
      • ACTL 6 antibody
      • ACTL 6A antibody
      • ACTL6 antibody
      • ACTL6A antibody
      • Arp4 antibody
      • ARPN BETA antibody
      • ArpNbeta antibody
      • BAF 53 antibody
      • BAF 53A antibody
      • BAF complex 53 kDa subunit antibody
      • BAF53 antibody
      • BRG1 associated factor 53A antibody
      • BRG1 associated factor antibody
      • BRG1/brm associated factor 53A antibody
      • hArpN beta antibody
      • INO80 complex subunit K antibody
      • INO80K antibody
      • MGC5382 antibody
      see all

    Anti-BAF53A antibody - ChIP Grade images

    • All lanes : Anti-BAF53A antibody - ChIP Grade (ab3882) at 1/1000 dilution

      Lane 1 : TEV elution of Tip60-TAP clone 1
      Lane 2 : TEV elution of Tip60-TAP clone 2
      Lane 3 : Total extract of Tip60-TAP clone 1
      Lane 4 : Total extract of Tip60-TAP clone 2


      Performed under reducing conditions.

      Predicted band size : 47 kDa

      This image is courtesy of Jacques Cote, Universite Laval, Quebec, Canada

      Total extracts from two different clones of MCF7 cells expressing a TAP tagged version of Tip60 were partially purified over IgG sepharose resin and eluted with TEV protease.  Western blot was performed using a 1/1000 dilution of the BAF53A antibody.

      Lane 1. TEV elution of Tip60-TAP clone #1
      Lane 2. TEV elution of Tip60-TAP clone #2
      Lane 3. Total extract of Tip60-TAP clone #1
      Lane 4. Total extract of Tip60-TAP clone #2

      NB: Tip60 is detected in the total extract due to the presence of Protein A in the TAP Tag (to which the primary and secondary abs bind). The TEV protease removes the Tag in lanes 1 and 2.

    • Sonicated Chromatin prepared from untreated (UI) or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure with ab3882 to BAF53A and the immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are nomalized over inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 8 µl of ab3882 and 2x106 cells were used in each ChIP experiment.

    • ab3882 at a 1/200 dilution staining asynchronous HeLa cells by ICC/IF. The cells were paraformaldehyde fixed and incubated with the antibody for 30 minutes at room temperature. Bound antibody was detected using a Cy3 conjugated Goat anti-rabbit antibody. Nuclei were visualised using DAPI stain. ab3882 gives a predominantly nuclear staining pattern.

      This image is courtesy of an Abreview submitted by Kirk McManus.

      See Abreview

    • All lanes : Anti-BAF53A antibody - ChIP Grade (ab3882) at 1 µg/ml

      Lane 1 : NIH 3T3 whole cell lysate (ab7179)
      Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
      Lane 3 : Mouse skeletal muscle tissue lysate - total protein (ab29711)
      Lane 4 : Ovary (Mouse) Tissue Lysate - normal tissue
      Lane 5 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size : 47 kDa
      Observed band size : 47 kDa
      Additional bands at : 120 kDa,18 kDa. We are unsure as to the identity of these extra bands.
    • Image courtesy of Human Protein Atlas
      ab3882 staining BAF53A in human ovary, showing a distinct and strong staining pattern of the nucleus of the ovarian stromal cells. Paraffin-embedded ovary tissue was incubated with ab3882 (1/35) for 30 minutes at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab3882 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

    References for Anti-BAF53A antibody - ChIP Grade (ab3882)

    This product has been referenced in:
    • Singh AP & Archer TK Analysis of the SWI/SNF chromatin-remodeling complex during early heart development and BAF250a repression cardiac gene transcription during P19 cell differentiation. Nucleic Acids Res 42:2958-75 (2014). IHC-P ; Mouse . Read more (PubMed: 24335282) »
    • Taulli R  et al. Failure to downregulate the BAF53a subunit of the SWI/SNF chromatin remodeling complex contributes to the differentiation block in rhabdomyosarcoma. Oncogene N/A:N/A (2013). Read more (PubMed: 23728344) »

    See all 5 Publications for this product

    Product Wall

    I have had the opportunity to inquire about ChIP testing for ab3882 and the image on our datasheet in particular. This image has been provided to us by a collaborator who tested this product in ChIP for us. This product is covered by our Abpromise ...

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    Thank you for contacting us. As discussed on phone we unfortunately do not sell small sizes of our products. The reason is that; we have over 70,000 products in our catalogue and having the small sized sample of each would be difficult to keep. How...

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    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Frozen sections)
    Sample Mouse Tissue sections (Embryonic Brain (E15))
    Specification Embryonic Brain (E15)
    Fixative Formaldehyde
    Permeabilization Yes - Triton 0.5%
    Blocking step BSA 0.1% + Normal Goat Serum 10% as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Sep 21 2007

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Specification HeLa
    Fixative Paraformaldehyde
    Username

    Dr. Kirk McManus

    Verified customer

    Submitted Jun 05 2006

    This antibody has been tested briefly in Immunofluorescence in U20S cells (human). IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"