Validated using a knockout cell line
Recombinant
RabMAb

Anti-BRG1 antibody [EPNCIR111A] (ab110641)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab110641 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 Use at an assay dependent concentration.
WB 1/10000 - 1/50000. Predicted molecular weight: 185 kDa.
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Antigen retrieval is recommended. Heat up to 98 °C, below boiling, and then let cool for 10-20 min.
ICC/IF 1/500.
Flow Cyt Use at an assay dependent concentration.

Target

  • FunctionTranscriptional coactivator cooperating with nuclear hormone receptors to potentiate transcriptional activation. Component of the CREST-BRG1 complex, a multiprotein complex that regulates promoter activation by orchestrating a calcium-dependent release of a repressor complex and a recruitment of an activator complex. In resting neurons, transcription of the c-FOS promoter is inhibited by BRG1-dependent recruitment of a phospho-RB1-HDAC repressor complex. Upon calcium influx, RB1 is dephosphorylated by calcineurin, which leads to release of the repressor complex. At the same time, there is increased recruitment of CREBBP to the promoter by a CREST-dependent mechanism, which leads to transcriptional activation. The CREST-BRG1 complex also binds to the NR2B promoter, and activity-dependent induction of NR2B expression involves a release of HDAC1 and recruitment of CREBBP. Belongs to the neural progenitors-specific chromatin remodeling complex (npBAF complex) and the neuron-specific chromatin remodeling complex (nBAF complex). During neural development a switch from a stem/progenitor to a post-mitotic chromatin remodeling mechanism occurs as neurons exit the cell cycle and become committed to their adult state. The transition from proliferating neural stem/progenitor cells to post-mitotic neurons requires a switch in subunit composition of the npBAF and nBAF complexes. As neural progenitors exit mitosis and differentiate into neurons, npBAF complexes which contain ACTL6A/BAF53A and PHF10/BAF45A, are exchanged for homologous alternative ACTL6B/BAF53B and DPF1/BAF45B or DPF3/BAF45C subunits in neuron-specific complexes (nBAF). The npBAF complex is essential for the self-renewal/proliferative capacity of the multipotent neural stem cells. The nBAF complex along with CREST plays a role regulating the activity of genes essential for dendrite growth. SMARCA4/BAF190A may promote neural stem cell self-renewal/proliferation by enhancing Notch-dependent proliferative signals, while concurrently making the neural stem cell insensitive to SHH-dependent differentiating cues (By similarity). Also involved in vitamin D-coupled transcription regulation via its association with the WINAC complex, a chromatin-remodeling complex recruited by vitamin D receptor (VDR), which is required for the ligand-bound VDR-mediated transrepression of the CYP27B1 gene. Acts as a corepressor of ZEB1 to regulate E-cadherin transcription and is required for induction of epithelial-mesenchymal transition (EMT) by ZEB1.
  • Tissue specificityColocalizes with ZEB1 in E-cadherin-negative cells from established lines, and stroma of normal colon as well as in de-differentiated epithelial cells at the invasion front of colorectal carcinomas (at protein level).
  • Involvement in diseaseDefects in SMARCA4 are the cause of rhabdoid tumor predisposition syndrome type 2 (RTPS2) [MIM:613325]. RTPS2 is a familial cancer syndrome predisposing to renal or extrarenal malignant rhabdoid tumors and to a variety of tumors of the central nervous system, including choroid plexus carcinoma, medulloblastoma, and central primitive neuroectodermal tumors. Rhabdoid tumors are the most aggressive and lethal malignancies occurring in early childhood.
  • Sequence similaritiesBelongs to the SNF2/RAD54 helicase family.
    Contains 1 bromo domain.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
    Contains 1 HSA domain.
  • Post-translational
    modifications
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATP dependent helicase SMARCA4 antibody
    • ATP-dependent helicase SMARCA4 antibody
    • BAF 190 antibody
    • BAF190 antibody
    • BAF190A antibody
    • Brahma protein like 1 antibody
    • BRG1 antibody
    • BRG1 associated factor 190A antibody
    • BRG1 protein antibody
    • BRG1-associated factor 190A antibody
    • BRM/SWI2 related gene 1 antibody
    • Global transcription activator homologous sequence antibody
    • global transcription activator snf2l4 antibody
    • Homeotic gene regulator antibody
    • hSNF2b antibody
    • Mitotic growth and transcription activator antibody
    • MRD16 antibody
    • Nuclear protein GRB1 antibody
    • Protein brahma homolog 1 antibody
    • Protein BRG-1 antibody
    • Protein BRG1 antibody
    • RTPS2 antibody
    • SMARC A4 antibody
    • SMARCA4 antibody
    • SMCA4_HUMAN antibody
    • SNF2 antibody
    • SNF2 beta antibody
    • SNF2 like 4 antibody
    • SNF2-beta antibody
    • SNF2B antibody
    • SNF2L4 antibody
    • SNF2LB antibody
    • Sucrose nonfermenting like 4 antibody
    • SWI/SNF related matrix associated actin dependent regulator of chromatin subfamily A member 4 antibody
    • SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4 antibody
    • SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 4 antibody
    • SWI2 antibody
    • Transcription activator BRG1 antibody
    see all

Anti-BRG1 antibody [EPNCIR111A] images



  • Predicted band size : 185 kDa
    Additional bands at : 185 kDa. We are unsure as to the identity of these extra bands.

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: BRG1 knockout HAP1 cell lysate (20 µg)
    Lane 3: K562 cell lysate (20 µg)
    Lane 4: Molt-4 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab110641 observed at 185 kDa. Red - loading control, ab18058, observed at 124 kDa.

    ab110641 was shown to specifically react with BRG1 when BRG1 knockout samples were used. Wild-type and BRG1 knockout samples were subjected to SDS-PAGE. ab110641 and ab18058 (loading control to Vinculin) were both diluted 1/10000 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 1 hour at room temperature before imaging.

  • ab110641 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 ab110641 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).

  • ab110641 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 ab110641 at 1/500 dilution and ab195889 (Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594)) at 1/250 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with ab150081 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)) 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).

  • ab110641 at 1/100 dilution staining BRG1 in Human testis tissue by Immunohistochemistry, Paraffin-embedded tissue.
  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling BRG1 with purified ab110641 at 1/200 dilution(10µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • All lanes : Anti-BRG1 antibody [EPNCIR111A] (ab110641) at 1/10000 dilution

    Lane 1 : K562 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : MOLT4 cell lysate
    Lane 4 : NIH3T3 cell lysate
    Lane 5 : PC12 cell lysate

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 185 kDa
  • ab110641 at 1/100 dilution staining BRG1 in Human kidney tissue by Immunohistochemistry, Paraffin-embedded tissue.
  • ChIP analysis using ab110641 binding BRG1 in mouse bone marrow derived macrophages. Cells were cross-linked for 10 minutes with formaldehyde. Samples were incubated with undiluted primary antibody for 16 hours at 4°C. Protein binding was detected using real-time PCR.
    Positive control: PU.1 antibody.
    Negative Control: rabbit IgG.

    See Abreview

References for Anti-BRG1 antibody [EPNCIR111A] (ab110641)

This product has been referenced in:
  • Su MT  et al. The SWI/SNF Chromatin Regulator, BRG1, Modulates the Transcriptional Regulatory Activity of the EBV DNA Polymerase Processivity Factor BMRF1. J Virol N/A:N/A (2017). Read more (PubMed: 28228591) »
  • Yoshida A  et al. Clinicopathological and molecular characterization of SMARCA4-deficient thoracic sarcomas with comparison to potentially related entities. Mod Pathol N/A:N/A (2017). IHC-P ; Human . Read more (PubMed: 28256572) »

See all 26 Publications for this product

Product Wall

Application Western blot
Sample Zebrafish Cell lysate - other (fibroblast cell tipe)
Gel Running Conditions Reduced Denaturing (SDS-PAGE 7.5%)
Loading amount 100 µg
Specification fibroblast cell tipe
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Jorge Castillo

Verified customer

Submitted Mar 28 2016

Application Western blot
Sample Mouse Tissue lysate - whole (Brain)
Gel Running Conditions Non-reduced Denaturing
Loading amount 50 µg
Specification Brain
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Jun 05 2015

Application ChIP
Detection step Real-time PCR
Sample Mouse Cell lysate - whole cell (bone marrow derived macrophages)
Specification bone marrow derived macrophages
Negative control rabbit IgG
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Positive control PU.1 antibody
Username

Dr. Silvia Bonifacio

Verified customer

Submitted May 02 2014

Application Immunoprecipitation
Immuno-precipitation step Protein G
Sample Human Cell lysate - nuclear (Human Cancer Cell line)
Specification Human Cancer Cell line
Total protein in input 500 µg
Username

Abcam user community

Verified customer

Submitted Apr 24 2014

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has c...

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I am sorry hear about the disappointing results. If customer have purchased the secondary antibody from Abcam (ab6718) and if the problem is due to secondary antibody then I am happy to replace it for this customer.
Please let me know the order num...

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Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.
I have read the details you have kindly provided and have following further questions for better understanding of the problem;
- What was ...

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Thank you for your enquiry.

At this time we have no data regarding the use of ab110641 in frozen sections. It certainly may be useful in this application, as many antibodies that are useful in IHC-P can also be used in frozen sections.
<...

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Thank you for providing details.
This is strange, if the red cells are seen even without primary antibody then the problem is either due to secondary antibody or due to protocol. I have checked the protocol and I find no problem with it other than ...

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