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  1. Link

    human-smarca4-brg1-knockout-hek-293t-cell-pellet-ab278821.pdf

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Neuroscience Cell Type Marker Neural Stem Cell marker
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Human SMARCA4 (BRG1) knockout HEK-293T cell pellet (ab278821)

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Western blot - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)
  • Western blot - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)
  • Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)
  • Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)

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Human UCHL1 (PGP9.5) knockout HEK-293T cell line (ab255443)

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Overview

  • Product name

    Human SMARCA4 (BRG1) knockout HEK-293T cell pellet
    See all BRG1 kits
  • Product overview

    Abcam’s knockout cell pellets give you access to native proteins, without the need to culture cells. Our knockout cell pellets are prepared from our single-gene knockout cell lines and provide an additional offering to our cell lysates.

    Cells are snap-frozen to provide high quality pellets that are suitable for extraction with alternative lysis buffers or for preparation of lysates from subcellular fractions. Our knockout cell pellets are suitable for a variety of applications, including PCR, gene expression profiling and DNA library preparation.

  • Parental Cell Line

    HEK293T
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 7 bp deletion in exon 4.
  • Passage number

    <20
  • Knockout validation

    Sanger Sequencing, Western Blot (WB)
  • Notes

    Pellet size: 5 million cells/vial.

    This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    Human SMARCA4 knockout HEK293T cell pellet 1 vial
    ab286211 - Human wild-type HEK293T cell pellet 1 vial
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Neural Stem Cell marker
    • Epigenetics and Nuclear Signaling
    • Chromatin Remodeling
    • Swi / Snf
    • Stem Cells
    • Signaling Pathways
    • Wnt
    • Nuclear
    • Epigenetics and Nuclear Signaling
    • Chromatin Remodeling
    • Trithorax
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • Other
  • Cell type

    epithelial
  • STR Analysis

    Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12

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.
  • Target information above from: UniProt accession P51532 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • ATP dependent helicase SMARCA4
    • ATP-dependent helicase SMARCA4
    • BAF 190
    • BAF190
    • BAF190A
    • Brahma protein like 1
    • BRG1
    • BRG1 associated factor 190A
    • BRG1 protein
    • BRG1-associated factor 190A
    • BRM/SWI2 related gene 1
    • Global transcription activator homologous sequence
    • global transcription activator snf2l4
    • Homeotic gene regulator
    • hSNF2b
    • Mitotic growth and transcription activator
    • MRD16
    • Nuclear protein GRB1
    • Protein brahma homolog 1
    • Protein BRG-1
    • Protein BRG1
    • RTPS2
    • SMARC A4
    • SMARCA4
    • SMCA4_HUMAN
    • SNF2
    • SNF2 beta
    • SNF2 like 4
    • SNF2-beta
    • SNF2B
    • SNF2L4
    • SNF2LB
    • Sucrose nonfermenting like 4
    • SWI/SNF related matrix associated actin dependent regulator of chromatin subfamily A member 4
    • SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4
    • SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 4
    • SWI2
    • Transcription activator BRG1
    see all

Associated products

  • KO cell lines

    • Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)
  • KO cell lysates

    • Human SMARCA4 (BRG1) knockout HEK-293T cell lysate (ab263853)
  • Related Products

    • Anti-BRG1 antibody [EPR3912] (ab108318)
    • Anti-BRG1 antibody [EPNCIR111A] (ab110641)
    • Anti-BRG1 antibody [EPNCIR111A] - BSA and Azide free (ab215998)
    • Anti-BRG1 antibody [EPR3912] - BSA and Azide free (ab222230)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab278821 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
WB
Use at an assay dependent concentration. Predicted molecular weight: 185 kDa.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 185 kDa.

Images

  • Western blot - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)
    Western blot - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)

    Lane 1: Wild-type HEK-293T cell lysate (20 ug)
    Lane 2: SMARCA4 knockout HEK-293T cell lysate (20 ug)

    Lanes 1- 2: Merged signal (red and green). Green - ab110641 observed at 185 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab110641 was shown to react with SMARCA4 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255432 (knockout cell lysate ab263853) was used. Wild-type HEK-293T and SMARCA4 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab110641 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 10000 dilution and a 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.

  • Western blot - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)
    Western blot - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)
    Lane 1: Wild-type HEK-293T cell lysate (20µg)

    Lane 2: SMARCA4 knockout HEK-293T cell lysate (20µg)

    Lanes 1- 2: Merged signal (red and green). Green - ab108318 observed at 185 kDa. Red - loading control ab130007 observed at 124 kDa.

    ab108318 Recombinant Anti-BRG1 antibody [EPR3912] was shown to specifically react with SMARCA4 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255432 (knockout cell lysate ab263853) was used. Wild-type and SMARCA4 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108318 and Anti-Vinculin antibody [VIN-54] 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.
  • Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)
    Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)

    Allele-1: 7 bp deletion in exon 4

     

  • Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)
    Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK293T cell pellet (ab278821)

    Allele-2: 1 bp deletion in exon 4

     

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (0)

Publishing research using ab278821? Please let us know so that we can cite the reference in this datasheet.

ab278821 has not yet been referenced specifically in any publications.

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