Overview

  • Product name
  • Description
    Rabbit polyclonal to BRG1
  • Host species
    Rabbit
  • Specificity
    We have conflicting data about the performance of this antibody in ChIP. Publications have reported positive results with this antibody using ChIP application, however we also have customer data indicating certain batches of this antibody did not work in ChIP in their hands. If you require any further information or assistance please contact Abcam Scientific Support Team.
  • Tested applications
    Suitable for: IP, WBmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1400 - 1500 of Human BRG1.

    Read Abcam's proprietary immunogen policy (Peptide available as ab13736.)

  • Positive control
    • This antibody gave a positive signal in the following cell lysate preparations: Jurkat; Jurkat nuclear; HepG2 nuclear.

Properties

Applications

Our Abpromise guarantee covers the use of ab4081 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
IP Use at an assay dependent concentration. PubMed: 20522713
WB 1/500 - 1/1000. Detects a band of approximately 235 kDa (predicted molecular weight: 185 kDa).
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function
      Transcriptional 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 specificity
      Colocalizes 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 disease
      Defects 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 similarities
      Belongs 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 localization
      Nucleus.
    • 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

    Images

    • Lane 1: Wild-type HAP1 cell lysate (40 µg)
      Lane 2: BRG1 knockout HAP1 cell lysate (40 µg)
      Lane 3: HeLa cell lysate (40 µg)
      Lane 4: K562 cell lysate (40 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab4081 observed at 240 kDa. Red - loading control, ab18058, observed at 124 kDa.

      ab4081 was shown to recognize BRG1 when BRG1 knockout samples were used, along with additional cross-reactive bands. Wild-type and BRG1 knockout samples were subjected to SDS-PAGE. Ab4081 and ab18058 (loading control to Vinculin) were diluted at 1/500 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) with 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.

    • All lanes : Anti-BRG1 antibody (ab4081) at 1 µg/ml

      Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
      Lane 2 : Jurkat nuclear extract lysate (ab14844)
      Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
      Lane 4 : HepG2 nuclear extract lysate (ab14660)

      Lysates/proteins at 10 µg per lane.

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

      Performed under reducing conditions.

      Predicted band size: 185 kDa
      Observed band size: 235 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 40 kDa. We are unsure as to the identity of these extra bands.

    • Rabbit polyclonal to BRG1 (ab4081) at 1/500 on HeLa Nuclear extract (20 ug per lane).

      Lane 1: ab4081 (1/500)
      Lane 2: ab4081 (1/500) + blocking peptide (1 ug)

      Secondary antibody: Goat anti-rabbit (HRP) - ab6721

    • Whole cell lysate prepared from HeLa cells was loaded at 150000 cells.
      ab4081 used at a 1/1000 dilution.
      The secondary used was an HRP conjugated donkey polyclonal used at a 1/5000 dilution.

      See Abreview

    References

    This product has been referenced in:
    • Del Gaudio N  et al. BRD9 binds cell type-specific chromatin regions regulating leukemic cell survival via STAT5 inhibition. Cell Death Dis 10:338 (2019). Read more (PubMed: 31000698) »
    • Gowda P  et al. Mutant Isocitrate Dehydrogenase 1 Disrupts PKM2-ß-Catenin-BRG1 Transcriptional Network-Driven CD47 Expression. Mol Cell Biol 38:N/A (2018). Read more (PubMed: 29463646) »
    See all 24 Publications for this product

    Customer reviews and Q&As

    21-26 of 26 Abreviews or Q&A

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Whole Mount Morula)
    Specification
    Whole Mount Morula
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.1% Triton-X 100
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted May 12 2011

    Application
    Western blot
    Sample
    Mouse Cell lysate - nuclear (Lung)
    Loading amount
    30 µg
    Specification
    Lung
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Jan 08 2008

    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (THP-1 cells (monocytes))
    Loading amount
    20 µg
    Specification
    THP-1 cells (monocytes)
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Nov 29 2007

    Answer

    I am really sorry ab4081 is proving difficult, I have spoken to our laboratory and asked to check that the current lot has not degraded, we are about to receive a new lot in a week or two and I would like to send you a vial of that lot as soon as it goes into stock. We would really appreciate if you could send us the image of your blots so we can compare them to our blots, to see what might be happening, I confirmed with the lab that the HeLa nuclear lysate was giving good signal (with a lower non specific band too though). Can you please also explain what you mean by "amplification"? Do you mean you got a signal? The antibody has so far only been tested on human cells so it is hard to work out if the problem with the rat and mouse samples is due to low cross reactivity, but it should definatly cross react with human. I look forward to receiving your images and to hear if you would like a replacement vial in a few weeks, I am happy to give you a credit note if you prefer not waiting, Thank you again for your patience,

    Read More

    Answer

    I'm sorry to hear you are having problem with ab4081. Thank you for taking the time to explain details of your experiment, it is very useful. The antibody actually recognises a 235kDa form of BRG1, which is what you have detected. You can see the blots of anti-BRG1 tested on nuclear extracts of HeLa cells on the datasheet of ab4081: the band detected was 235kDa in size and the use of a blocking peptide could abolish this signal, confirming the antibody is specific for BRG1. Although the predicted molecular weight of BRG1 is 185kDa, the protein has numerous modifications sites which can explain its higher molecular weight: BRG1 has a site for a poly-Lys chain extension, several poly-Glu extension sites, a Bromodomain and a HSA domain (human serum albumin domain). I would recommend using nuclear extraction rather than whole lysates, BSA blocking (1hr 5%v/v in TBST) and diluting the primary and secondary antibodies in TBST only. Please let me know if this helps and do not hesitate to contact us for further advice,

    Read More

    Question
    Answer

    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% milk / 0.2% tween for one hour. Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes. All blocking and incubation steps carried out at 37 degrees. No staining was observed. However, staining was only attempted once and this is by no means an indication that the antibody doesn't work in IF, it merely did not perform under the conditions above. Please see reviews and the datasheet for details of any further testing.

    Read More

    21-26 of 26 Abreviews or Q&A

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