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:
    • Pandolfini L  et al. RISC-mediated control of selected chromatin regulators stabilizes ground state pluripotency of mouse embryonic stem cells. Genome Biol 17:94 (2016). Read more (PubMed: 27154007) »
    • Mita P  et al. URI Regulates KAP1 Phosphorylation and Transcriptional Repression via PP2A Phosphatase in Prostate Cancer Cells. J Biol Chem 291:25516-25528 (2016). Read more (PubMed: 27780869) »
    See all 21 Publications for this product

    Customer reviews and Q&As

    1-10 of 26 Abreviews or Q&A

    Answer

    Vielen Dank für Ihren Anruf.

    Es tut mir leid, dass Sie Probleme mit zwei Röhrchen dieses Antikörpers hatten. Wie am Telefon besprochen, habewir eine kostenlose Ersatzlieferung mit einem Röhrchen von derneuen Charge des ab4081 für Sie in Auftrag gegeben. Sie hat die Referenznummer xx und sollte morgen bei Ihnen ankommen.

    Bitte lassen Sie uns wissen, ob die neue Charge in ChIP funktioniert.Ichfreue mich wieder von Ihnen zu hören.

    Read More

    Question
    Answer

    DISCOUNT CODE: ******
    Expiration date: ********

    I am very pleased to hear you would like to accept our offer and test ab118558 inXenopus laevis. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for Xenopus laevis and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


    Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

    Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

    The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

    Read More
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U2OS)
    Permeabilization
    Yes - NP-40
    Specification
    U2OS
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted May 16 2016

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    ChIP
    Sample
    Mouse Cell lysate - other (...)
    Specification
    ...
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: formaldehyde
    Detection step
    Real-time PCR
    Positive control
    The ChIP experiment with this antibody was performed side by side with an antibody that recognizes another subunit of the complex. That antibody gave a good signal, around 1-3%IP, whereas antibody ab4081 had a very low signal 0.01-0.02%IP, and therefore the numbers were deemed unreliable.
    Negative control
    A rabbit IgG IP control was performed in the same experiment and had numbers comparable to the ab4081 ChIP (around 0.01%).

    Abcam user community

    Verified customer

    Submitted Nov 07 2012

    Question

    I've looked at the papers and it seems very hard to know what is going on. The data in Kia et al., 2008 looks good since they do show the %IP (around 0.2 to 0.4), which seem to be reasonable numbers. They also have good controls etc. The Bedadala paper is hard to judge, since there is only one figure (Fig. 5A), where a band is there upon induction but not without. It is hard to know what the %IP is, since they don't quantify their ChIP. Kia was done in human cells, and Bedadala in mouse, I did my ChIP in mouse. Maybe it works better in human cells……

    I'm not sure you are familiar with ChIP but the problem is often how to know whether there is a signal or not. IgG controls are almost always lower, and unless you have a positive control or a knockdown where the signal disappears it is very hard to judge. However, antibodies are coming under more and more scrutiny and as a provider of high grade antibodies it must be in your interest to serve the community. I have used your histone, and histone modification antibodies in the past and they seem to work fine.

    It would be very helpful, if you indicated in your antibody descriptions where the data came from so that one could easily see, whether it came from in house experiments or from the literature. I did not find that information easily, although it probably would not have helped either. And I would feel much more comfortable if AbCam performed ChIP verification themselves, before an antibody was advertised as ChIP-grade. Otherwise the scientific community does the QC and pays for it too.

    Read More
    Answer

    Thanks so much for your email. I will certainly pass your comments along. In addition, please note that your comments will appear under the 'Scientific Support' tab on the online product datasheet for ab4081.

    I hope you were able to submit your review successfully. Please contact me if you have any further comments or questions.

    Read More

    Answer

    The antibody being noted as ChiP-tested is based on two publications on our 'Specific references' tab on the product datasheet:

    1. Bedadala GR et al. Thyroid hormone controls the gene expression of HSV-1 LAT and ICP0 in neuronal cells. Cell Res 20:587-98 (2010).
    ChIP. PubMed: 20386570
    2. Kia SK et al. SWI/SNF mediates polycomb eviction and epigenetic reprogramming of the INK4b-ARF-INK4a locus. Mol Cell Biol 28:3457-64
    (2008). WB, ChIP; Human. PubMed: 18332116
    I hope this helps. Please contact me again if you have any further questions.

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Apr 18 2012

    Answer

    Thank you for contacting us.

    ab4081 is predicted to react with Rat so can certainly give you the discount code for this ab. ab70558 is also eligible for discount collaboration.

    Please note these antibodies are not phospho specific so could you explain how you will be using these antibodies for identification of phosphorylated BRG1?

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (HeLa cells)
    Specification
    HeLa cells
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: Formaldehyde
    Detection step
    Real-time PCR
    Positive control
    I examined several cites that were reported to be occupied by BRG1, such as p53 and p21 promoters, CD44 transcriptional start site and transcribed region

    Dr. Efrat Shema

    Verified customer

    Submitted Apr 04 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa cells)
    Loading amount
    20 µg
    Specification
    HeLa cells
    Treatment
    siRNA transfections 40 hours
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 40 minute(s) · Concentration: 4% · Temperature: 25°C

    Dr. Efrat Shema

    Verified customer

    Submitted Apr 04 2012

    1-10 of 26 Abreviews or Q&A

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