Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3912] to BRG1 - BSA and Azide free
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-BRG1 antibody [EPR3912] - BSA and Azide free
    See all BRG1 primary antibodies

  • Description

    Rabbit monoclonal [EPR3912] to BRG1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Rat
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HAP1, HEK-293, K562, and Molt-4 cell lysate. ICC/IF: HeLa cells. IHC-P: colon tissue. Flow Cyt: HeLa cells.

  • General notes

    ab222230 is the carrier-free version of ab108318. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab222230 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Western blot - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    All lanes : Anti-BRG1 antibody [EPR3912] (ab108318) at 1/10000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : SMARCA4 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 185 kDa
    Observed band size: 185 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab108318).

      Lanes 1- 2: Merged signal (red and green). Green - ab108318 observed at 185 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

     ab108318 was shown to react with BRG1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255432 (knockout cell lysate ab263853) was used. Wild-type HEK-293T and SMARCA4 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108318 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder cancer tissue sections labeling BRG1 with purified ab108318 at 1/500 dilution (0.23 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

  • Western blot - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Western blot - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    All lanes : Anti-BRG1 antibody [EPR3912] (ab108318) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : BRG1 knockout HAP1 cell lysate
    Lane 3 : K562 cell lysate
    Lane 4 : Molt-4 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 185 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab108318).

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

    ab108318 was shown to specifically react with BRG1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when BRG1 knockout samples were used. Wild-type and BRG1 knockout samples were subjected to SDS-PAGE, ab108318 and ab18058 (loading control to Vinculin) were diluted 1/1000 and 1/10,000 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 1/10,000 dilution for 1hr at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling BRG1 with purified ab108318 at 1/50 dilution (2.3 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) at 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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

  • Flow Cytometry - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Flow Cytometry - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling BRG1 with purified ab108318 at 1/20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat stomach tissue sections labeling BRG1 with purified ab108318 at 1:500 dilution (0.23 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). 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.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse stomach tissue sections labeling BRG1 with purified ab108318 at 1/500 dilution (0.23 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

  • Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    ab108318 (unpurified) staining BRG1 in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab108318 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

  • Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    ab108318 (unpurified) staining BRG1 in wild-type HAP1 cells (top panel) and BRG1 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab108318 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    ab108318 (unpurified) at 1/100 staining BRG1 in paraffin-embedded Human colon tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    ab108318 (unpurified) showing positive staining in Urinary bladder transitional carcinoma tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    ab108318 (unpurified) showing positive staining in Breast carcinoma tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    ab108318 (unpurified) showing positive staining in Ovarian carcinoma tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

    ab108318 (unpurified) showing positive staining in Normal tonsil tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108318).

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)
    Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

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