Overview

  • Product name

    Anti-BANF1/BAF antibody [EPR7668]
    See all BANF1/BAF primary antibodies
  • Description

    Rabbit monoclonal [EPR7668] to BANF1/BAF
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Dog, Human
  • Immunogen

    Synthetic peptide within Human BANF1/BAF. The exact sequence is proprietary.
    Database link: O75531

  • Positive control

    • WB: HeLa, HepG2, MDCK, and 293T cell lysates. IHC-P: Human colonic carcinoma, and Human ovarian carcinoma tissues. ICC/IF: HeLa, HepG2 and NIH/3T3 cells. Flow Cyt: HeLa cells.
  • General notes

     

     

     This product was previously labelled as BANF1

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Dissociation constant (KD)

    KD = 2.18 x 10 -10 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR7668
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab129184 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
WB 1/1000 - 1/10000. Predicted molecular weight: 10 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt 1/80.

For unpurified use at 1/200

ICC/IF 1/100.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Plays fundamental roles in nuclear assembly, chromatin organization, gene expression and gonad development. May potently compress chromatin structure and be involved in membrane recruitment and chromatin decondensation during nuclear assembly. Contains 2 non-specific dsDNA-binding sites which may promote DNA cross-bridging. Exploited by retroviruses for inhibiting self-destructing autointegration of retroviral DNA, thereby promoting integration of viral DNA into the host chromosome. EMD and BAF are cooperative cofactors of HIV-1 infection. Association of EMD with the viral DNA requires the presence of BAF and viral integrase. The association of viral DNA with chromatin requires the presence of BAF and EMD.
    • Tissue specificity

      Widely expressed. Expressed in colon, brain, heart, kidney, liver, lung, ovary, pancreas, placenta, prostate, skeletal muscle, small intestine, spleen and testis. Not detected in thymus and peripheral blood leukocytes.
    • Sequence similarities

      Belongs to the BAF family.
    • Domain

      Has a helix-hairpin-helix (HhH) structural motif conserved among proteins that bind non-specifically to DNA.
      LEM domain proteins bind centrally on the BAF dimer, whereas DNA binds to the left and right sides.
    • Post-translational
      modifications

      Partially phosphorylated on serine. Ser-4 phosphorylation may block BAF ability to promote EMD binding to lamins in vitro. Non phosphorylated BAF seems to enhances binding between EMD and LMNA.
    • Cellular localization

      Nucleus. Cytoplasm. Chromosome. Significantly enriched at the nuclear inner membrane, diffusely throughout the nucleus during interphase and concentrated at the chromosomes during the M-phase. May be included in HIV-1 virions via its interaction with viral GAG polyprotein.
    • Information by UniProt
    • Database links

    • Alternative names

      • BAF antibody
      • BAF_HUMAN antibody
      • BANF 1 antibody
      • BANF1 antibody
      • Barrier to autointegration factor 1 antibody
      • Barrier to autointegration factor antibody
      • Barrier-to-autointegration factor antibody
      • BCRG 1 antibody
      • BCRG1 antibody
      • BCRP 1 antibody
      • BCRP1 antibody
      • Breakpoint cluster region protein 1 antibody
      • D14S1460 antibody
      • MGC111161 antibody
      • NGPS antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human ovarian carcinoma tissue sections labeling BANF1/BAF with Purified ab129184 at 1:100 dilution (0.78 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    • Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling BANF1/BAF with Purified ab129184 at 1:100 dilution (7.8 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • All lanes : Anti-BANF1/BAF antibody [EPR7668] (ab129184) at 1/1000 dilution

      Lane 1 : HeLa cell lysates
      Lane 2 : HepG2 cell lysates
      Lane 3 : 293T cell lysates

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 10 kDa

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryo fibroblast cells)cells labeling BANF1/BAF with ab129184 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing mainly nuclear with cytoplasmic staining on NIH/3T3 cell line. The nuclear counterstain is DAPI (blue).

    • ab129184 staining BANF1/BAF in MDCK cell pellets in paraffin by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Cell pellets were fixed with paraformaldehyde, permeabilized with Tween-20 and blocked with 1% serum for 2 hours at room temperature; antigen retrieval was by heat mediation in Tris/EDTA pH 9. Samples were incubated with primary antibody (1/100 in 1% BSA + 1% FBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

      See Abreview

    • Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling BANF1/BAF (red) with ab129184 at a 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with primary and secondary antibodies.

    • ab129184, at a 1/100 dilution, staining BANF1/BAF in paraffin embedded Human colonic carcinoma tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma) cells labeling BANF1/BAF with ab129184 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing mainly nuclear with weakly cytoplasmic staining on Hela cell line. The nuclear counterstain is DAPI (blue).

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    • Birendra Kc  et al. VRK2A is an A-type lamin-dependent nuclear envelope kinase that phosphorylates BAF. Mol Biol Cell 28:2241-2250 (2017). Read more (PubMed: 28637768) »
    See 1 Publication for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (ISolated embryonic cardiomyocytes at stage E16.5)
    Permeabilization
    Yes - 0.2% triton X100/PBS
    Specification
    ISolated embryonic cardiomyocytes at stage E16.5
    Blocking step
    (agent) for 30 minute(s) · Concentration: 5% · Temperature: 20°C
    Fixative
    Paraformaldehyde

    Jacob Ross

    Verified customer

    Submitted Jun 21 2019

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (NMuMG)
    Permeabilization
    Yes - Triton 0,1%
    Specification
    NMuMG
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jan 29 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue sections (MDCK pellets in paraffin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9
    Permeabilization
    Yes - Tween-20
    Specification
    MDCK pellets in paraffin
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 03 2014

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (mouse pancreas tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH6
    Permeabilization
    Yes - Tween-20
    Specification
    mouse pancreas tumor
    Blocking step
    BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Nov 25 2014

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