Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)

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Overview

  • Product name

    Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free
    See all BRG1 primary antibodies

  • Description

    Rabbit monoclonal [EPNCIR111A] to BRG1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, Flow Cyt, ICC/IF, IP, ChIPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Mouse BRG1 aa 200-300.

  • Positive control

    • WB: HEK-293, K562, HeLa, MOLT4, NIH3T3, and PC12 cell lysates. IHC-P: Human kidney and testis tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.

  • General notes

    ab215998 is the carrier-free version of ab110641 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab215998 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab215998 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Heat up to 98°C, below boiling, and then let cool for 10-20 min.
WB Use at an assay dependent concentration. Predicted molecular weight: 185 kDa.
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ChIP 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

    see all

  • 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

  • Western blot - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)
    Western blot - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)
    All lanes : Anti-BRG1 antibody [EPNCIR111A] (ab110641) at 1/10000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : SMARCA4 knockout HAP1 cell lysate
    Lane 3 : Wild-type HEK-293T cell lysate
    Lane 4 : SMARCA4 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 185 kDa
    Observed band size: 185 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab110641).

    Lanes 1-4: Merged signal (red and green). Green - ab110641 observed at 185 kDa. Red - loading control, ab130007 observed at 125 kDa.

    ab110641 was shown to react with BRG1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout sample ab263853 was used. Wild-type and BRG1 knockout samples were subjected to SDS-PAGE. ab110641 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • ChIP - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)
    ChIP - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)

    This ChIP data was generated using the same anti-BRG1 antibody clone, EPNCIR111A, in a different buffer formulation (cat# ab110641).

    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 undiluted 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)
    Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab110641).

  • Flow Cytometry - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)
    Flow Cytometry - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab110641).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)

    ab110641 at 1/100 dilution staining BRG1 in Human kidney tissue by Immunohistochemistry, Paraffin-embedded tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab110641).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)
    Immunocytochemistry/ Immunofluorescence - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)

    This ICC/IF data was generated using the same anti-BRG1 antibody clone, EPNCIR111A, in a different buffer formulation (cat# ab110641).

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)

    This IHC data was generated using the same anti-BRG1 antibody clone, EPNCIR111A, in a different buffer formulation (cat# ab110641).

    ab110641 at 1/100 dilution staining BRG1 in Human testis tissue by Immunohistochemistry, Paraffin-embedded tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Otte A  et al. In vitro and in vivo therapeutic approach for a small cell carcinoma of the ovary hypercalcaemic type using a SCCOHT-1 cellular model. Orphanet J Rare Dis 9:126 (2014). WB ; Human . Read more (PubMed: 25103190) »
  • Goljanek-Whysall K  et al. myomiR-dependent switching of BAF60 variant incorporation into Brg1 chromatin remodeling complexes during embryo myogenesis. Development 141:3378-87 (2014). WB ; Mouse . Read more (PubMed: 25078649) »
See all 4 Publications for this product

Publishing research using ab215998? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

ChIP abreview for Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free

 Average
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (MCF7 breast cancer cell line and tumour samples)
Negative control
IgG was used as a negative control and calculated relative abundance of BRG1 peptides in mass spectrometry.
Specification
MCF7 breast cancer cell line and tumour samples
Detection step
Other
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 30 minute(s) and 0 second(s)
Specification of the cross-linking agent: DSG (30 mins) + Formaldehyde (
Positive control
H3K27ac was used as a positive control
Read More

Dr. Sankari Nagarajan

Verified customer

Submitted Apr 03 2019

ChIP abreview for Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free

 Good
Abreviews
abreview image
Application
ChIP
Sample
Mouse Cell lysate - nuclear (primary bone marrow derived macrophages)
Negative control
IgG antibody
Specification
primary bone marrow derived macrophages
Detection step
Other
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
ChIP was performed in parallel to H3K4me1, as positive control
Read More

Abcam user community

Verified customer

Submitted Jun 28 2018

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