概述

  • 产品名称Anti-BRG1抗体
    参阅全部 BRG1 一抗
  • 描述
    兔多克隆抗体to BRG1
  • 特异性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.
  • 经测试应用适用于: IP, WB, ICC/IFmore details
    不适用于: IHC-P
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Rat
  • 免疫原

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

    (Peptide available as ab13736.)

  • 阳性对照
    • This antibody gave a positive signal in the following cell lysate preparations: Jurkat; Jurkat nuclear; HepG2 nuclear.

性能

应用

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.

应用 Ab评论 说明
IP Use at an assay dependent dilution. PubMed: 20522713
WB 1/500 - 1/1000. Detects a band of approximately 235 kDa (predicted molecular weight: 185 kDa).
ICC/IF 1/200.
  • 应用说明Is unsuitable for IHC-P.
  • 靶标

    • 功能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.
    • 组织特异性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).
    • 疾病相关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.
    • 序列相似性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.
    • 翻译后修饰Phosphorylated upon DNA damage, probably by ATM or ATR.
    • 细胞定位Nucleus.
    • Information by UniProt
    • 数据库链接
    • 别名
      • 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 图像



    • Predicted band size : 185 kDa

      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) ab216773 with Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.



    • Predicted band size : 185 kDa

      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

    • 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
      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.

    • Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 185 kDa


      Exposure time : 8 minutes

    • Developed using the ECL technique

      Predicted band size : 185 kDa
      Observed band size : 225 kDa (why is the actual band size different from the predicted?)


      Exposure time : 5 minutes

      This image is courtesy of an anonymous abreview.

      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

    • ab4081 (1/200) staining BRG1 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised in 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

      See Abreview

    Anti-BRG1 antibody (ab4081)参考文献

    This product has been referenced in:
    • Sexton BS  et al. The spring-loaded genome: nucleosome redistributions are widespread, transient, and DNA-directed. Genome Res 24:251-9 (2014). WB . Read more (PubMed: 24310001) »
    • Maida Y  et al. Involvement of telomerase reverse transcriptase in heterochromatin maintenance. Mol Cell Biol 34:1576-93 (2014). IP . Read more (PubMed: 24550003) »

    See all 13 Publications for this product

    Product Wall

    Vielen Dank für Ihren Anruf.

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    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 ...

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    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
    Username

    Abcam user community

    Verified customer

    提交于 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%).
    Username

    Abcam user community

    Verified customer

    提交于 Nov 07 2012

    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 yo...

    Read More

    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 ...

    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
    Username

    Dr. Kirk McManus

    Verified customer

    提交于 Apr 18 2012

    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 ...

    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
    Username

    Dr. Efrat Shema

    Verified customer

    提交于 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
    Username

    Dr. Efrat Shema

    Verified customer

    提交于 Apr 04 2012

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