遺伝子ノックアウト細胞株 検証済

Anti-BRG1 抗体 (ab4081)

Reviews (10)Q&A (16)References (26)
Western blot - Anti-BRG1 antibody (ab4081)
  • Western blot - Anti-BRG1 antibody (ab4081)
  • Western blot - Anti-BRG1 antibody (ab4081)
  • Western blot - Anti-BRG1 antibody (ab4081)
  • Western blot - Anti-BRG1 antibody (ab4081)

Key features and details

  • Rabbit polyclonal to BRG1
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-BRG1 antibody [EPNCIR111A] (ab110641)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-BRG1 antibody
    BRG1 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to BRG1

  • 由来種

    Rabbit

  • 特異性

    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.

  • アプリケーション

    適用あり: WBmore details
    適用なし: IHC-P

  • 種交差性

    交差種: Human, Recombinant fragment
    交差が予測される動物種: Mouse, Rat

  • 免疫原

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

  • 特記事項

    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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab4081の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB (4)
1/500 - 1/1000. Detects a band of approximately 235 kDa (predicted molecular weight: 185 kDa).
特記事項
WB
1/500 - 1/1000. Detects a band of approximately 235 kDa (predicted molecular weight: 185 kDa).
追加情報
Is unsuitable for IHC-P.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    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).

  • 関連疾患

    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.

  • 配列類似性

    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.

  • 翻訳後修飾

    Phosphorylated upon DNA damage, probably by ATM or ATR.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • 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

画像

  • Western blot - Anti-BRG1 antibody (ab4081)
    Western blot - Anti-BRG1 antibody (ab4081)
    All lanes : Anti-BRG1 antibody (ab4081) at 1/500 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : BRG1 knockout HAP1 cell lysate
    Lane 3 : HeLa (Human epithelial adenocarcinoma cell line) whole cell lysate
    Lane 4 : K562 (Human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 185 kDa



    Lanes 1 - 4: Merged signal (red and green).

    Green - ab4081 observed at 240 kDa.

    Red - loading control, Mouse monoclonal Anti-Vinculin antibody, 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 loading control to Vinculin were diluted at 1/500 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773 - Green) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776 - Red) secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-BRG1 antibody (ab4081)
    Western blot - Anti-BRG1 antibody (ab4081)
    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
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Hep G2 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.

  • Western blot - Anti-BRG1 antibody (ab4081)
    Western blot - Anti-BRG1 antibody (ab4081)
    All lanes : Anti-BRG1 antibody (ab4081) at 1/500 dilution

    Lane 1 : HeLa (Human epithelial adenocarcinoma cell line) Nuclear extract
    Lane 2 : HeLa Nuclear extract with Human BRG1 peptide (ab13736) at 1 µg

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) (Goat anti-rabbit (HRP))

    Predicted band size: 185 kDa

  • Western blot - Anti-BRG1 antibody (ab4081)
    Western blot - Anti-BRG1 antibody (ab4081)
    Anti-BRG1 antibody (ab4081) at 1/1000 dilution + Recombinant human BRG1 protein (ab82237) at 0.1 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 185 kDa


    Exposure time: 8 minutes
  • Western blot - Anti-BRG1 antibody (ab4081)
    Western blot - Anti-BRG1 antibody (ab4081)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

参考文献 (26)

ab4081 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab4081 は 26 報の論文で使用されています。

  • 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). PubMed: 31000698
  • Lei I  et al. SWI/SNF Component BAF250a Coordinates OCT4 and WNT Signaling Pathway to Control Cardiac Lineage Differentiation. Front Cell Dev Biol 7:358 (2019). PubMed: 32039194
  • Wang J  et al. Asymmetric Expression of LincGET Biases Cell Fate in Two-Cell Mouse Embryos. Cell 175:1887-1901.e18 (2018). PubMed: 30550787
  • 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). PubMed: 29463646
  • Khan AW  et al. MeCP2 interacts with chromosomal microRNAs in brain. Epigenetics 12:1028-1037 (2017). PubMed: 29412786
View all Publications for this product

レビューと Q&A

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レビューを送る 質問を送る

1-10 of 26 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-BRG1 antibody

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

投稿 May 16 2016

ChIP abreview for Anti-BRG1 antibody

 Poor
Abreviews
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%).
Read More

Abcam user community

Verified customer

投稿 Nov 07 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-BRG1 antibody

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X100 in PBS
Read More

DR. Kirk Mcmanus

Verified customer

投稿 Apr 18 2012

ChIP abreview for Anti-BRG1 antibody

 Poor
Abreviews
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
Read More

Dr. Efrat Shema

Verified customer

投稿 Apr 04 2012

Western blot abreview for Anti-BRG1 antibody

 Excellent
Abreviews
abreview image
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
Read More

Dr. Efrat Shema

Verified customer

投稿 Apr 04 2012

Question

Phone call: Batch xx und Batch xx tested with 6 ug/ml and 8 ug/ml. This did not improve the signal on same samples (F16 cells). Send a vial of new batch

Read More
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

We would like to test ab118558 in Xenopus.
Thanks,

Read More
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.

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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.

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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

Question

Its noted this antibody has been tested successfully in ChIP. Is there any ChIP data available for this antibody?

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.

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Question

Hello,
I would like to useAnti-BRG1 antibody (ab4081), andAnti-BRG1 antibody (ab70558) in rat. Additionally I would like to test them for recognizing Phosphorylated forms of BRG1. May I become a part of your Testing discount program. No information exists for rat or for identification of phosphorylated BRG1 from these Antibodies. I think that both experiments would be useful for your reviews.
I can order these Antibodies today if that works for you.

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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

1-10 of 26 Abreviews or Q&A

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