Overview

  • Product name
    Anti-BRG1 antibody [EPNCIR111A]
    See all BRG1 primary antibodies
  • Description
    Rabbit monoclonal [EPNCIR111A] to BRG1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ChIP, WB, IP, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Mouse BRG1 aa 200-300. The exact sequence is proprietary.
    Database link: P51532

  • Positive control
    • K562, HeLa, MOLT4, NIH3T3, and PC12 cell lysates; Human kidney and testis tissues; HeLa cells. ICC/IF: HeLa cells, SMARCA4-HAP1 cells
  • General notes

    This antibody was developed as part of a collaboration between Epitomics, the National Cancer Institute's Center for Cancer Research and the lab of Gordon Hager. View antibodies from NCI Center for Cancer Research Collaboration.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

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.Can be blocked with BRG1 peptide (ab241115).
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

  • 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 (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 in wild-type HAP1 cells. No band was observed 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 1/10,000 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 1hr at room temperature before imaging.

  • Downregulation of BAF47 alters muscle terminal differentiation and cell cycle exit

    BAF47 and BRG1 occupancy at cyclin D1 promoter. ChIP-qPCR analyses of BAF47 and BRG1 in myoblast in proliferating C2C12 cells and at 24 h of differentiation. The immunoprecipitated material was quantified by qPCR, and results are expressed as fold enrichment of the % of Input of BAF47 or BRG1 ChIP over % of Input of the IgG average. Data are represented as mean ±SEM, n = 3.

    BRG1 was percipiteted using ab110641 overnight at 4°C.

    (From Figure 3A of Joliot et al)

  • 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).

  • 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 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

  • 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.

  • 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).

  • 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.

References

This product has been referenced in:
  • Shao J  et al. Angiotensin II induced CSF1 transcription is mediated by a crosstalk between different epigenetic factors in vascular endothelial cells. Biochim Biophys Acta Gene Regul Mech 1862:1-11 (2019). Read more (PubMed: 30317027) »
  • Xue Y  et al. SMARCA4 loss is synthetic lethal with CDK4/6 inhibition in non-small cell lung cancer. Nat Commun 10:557 (2019). Read more (PubMed: 30718506) »
See all 58 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

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

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

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

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

Abcam user community

Verified customer

Submitted Apr 24 2014

Answer

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 caused. As requested, I have issued a free of charge replacement with the order number 1231536.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Read More

Answer

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 number for secondary antibody.

Read More

Answer

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 the cell line used? e.g. human liver cancer cell line.
- Do you see difference in no primary control? Were the cells red in colour as seen with primary antibody?
- Do you think the problem is due to ab6718 or ab110641.
- What was the storage condition of antibody?
- Did you change any step when doing repeat experiment?
Thank you very much for your cooperation. I will look forward to hearing from you soon.

Read More

Answer

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.

Here is a link to a general IHC-Fr protocol on our website that you may find useful:
https://www.abcam.com/index.html?pageconfig=resource&rid=11383

I hope this helps. Please contact me again if you have any further questions.

Read More

Answer

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 the wash step which I assume is 3 washes of 5 minutes.
You may need to try again, following the correct ICC/IF protocol steps.
Please let me know if the results do not improve.

Read More

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