Overview

  • Product name

    Anti-Baf180 antibody [EPR15860]
    See all Baf180 primary antibodies
  • Description

    Rabbit monoclonal [EPR15860] to Baf180
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Baf180 aa 500-700. The exact sequence is proprietary.
    Database link: Q86U86

  • Positive control

    • WB: Human fetal kidney lysate; HeLa nuclear lysate; IHC-P: Human kidney tissue; ICC/IF: HeLa cells.
  • General notes

     

     

    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

Applications

Our Abpromise guarantee covers the use of ab196022 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 Use at an assay dependent concentration.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/10000. Detects a band of approximately 193 kDa (predicted molecular weight: 193 kDa).
ICC/IF 1/100.

Target

  • Function

    Involved in transcriptional activation and repression of select genes by chromatin remodeling (alteration of DNA-nucleosome topology). Act as a negative regulator of cell proliferation.
  • Tissue specificity

    Widely expressed.
  • Involvement in disease

    Defects in PBRM1 are a cause of renal cell carcinoma (RCC) [MIM:144700]. It is a heterogeneous group of sporadic or hereditary carcinoma derived from cells of the proximal renal tubular epithelium. It is subclassified into clear cell renal carcinoma (non-papillary carcinoma), papillary renal cell carcinoma, chromophobe renal cell carcinoma, collecting duct carcinoma with medullary carcinoma of the kidney, and unclassified renal cell carcinoma.
  • Sequence similarities

    Contains 2 BAH domains.
    Contains 6 bromo domains.
    Contains 1 HMG box DNA-binding domain.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • BAF180 antibody
    • BRG1-associated factor 180 antibody
    • CG11375 antibody
    • hPB1 antibody
    • MGC156155 antibody
    • MGC156156 antibody
    • OTTHUMP00000202149 antibody
    • OTTHUMP00000202150 antibody
    • OTTHUMP00000202151 antibody
    • OTTHUMP00000202153 antibody
    • OTTHUMP00000202163 antibody
    • PB1 antibody
    • PB1_HUMAN antibody
    • Pbrm1 antibody
    • polybromo 1 antibody
    • Polybromo-1D antibody
    • Protein polybromo-1 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Baf180 knockout HAP1 cell lysate (20 µg)
    Lane 3: Human kidney tissue lysate (20 µg)
    Lane 4: HeLa cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab196022 observed at 193 kDa. Red - loading control, ab18058, observed at 124 kDa.

    ab196022 was shown to recognize Baf180 when Baf180 knockout samples were used, along with additional cross-reactive bands. Wild-type and Baf180 knockout samples were subjected to SDS-PAGE. ab196022 and ab18058 (loading control to Vinculin) were diluted 1/5000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • ab196022 staining Baf180 in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. The sample was incubated with the primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • Anti-Baf180 antibody [EPR15860] (ab196022) at 1/10000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 193 kDa
    Observed band size: 193,35 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    5% NFDM/TBST: Blocking and diluting buffer.

     

  • Anti-Baf180 antibody [EPR15860] (ab196022) at 1/10000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) nuclear lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 193 kDa
    Observed band size: 193 kDa


    Exposure time: 15 seconds


    5% NFDM/TBST: Blocking and diluting buffer.

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Baf180 using ab196022 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human kidney tissue is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

     

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Baf180 with ab196022 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). The nuclear counter stain is DAPI (blue).

    Nuclear staining on HeLa cell line is observed.

    -ve control -  Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution. The nuclear counter stain is DAPI (blue).

     

References

This product has been referenced in:

  • Cabot B  et al. Differential expression of key subunits of SWI/SNF chromatin remodeling complexes in porcine embryos derived in vitro or in vivo. Mol Reprod Dev 84:1238-1249 (2017). ICC/IF . Read more (PubMed: 29024220) »
See 1 Publication for this product

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