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  1. Link

    smarca2--brm-antibody-epr23103-44-ab240648.pdf

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Epigenetics and Nuclear Signaling Chromatin Remodeling Swi / Snf
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)

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  • SDS
  • Certificate of Compliance
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Western blot - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
  • Western blot - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
  • Flow Cytometry (Intracellular) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
  • Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
  • Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
  • Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23103-44] to SMARCA2 / BRM
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 APC Carrier Free HRP PE

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Overview

  • Product name

    Anti-SMARCA2 / BRM antibody [EPR23103-44]
    See all SMARCA2 / BRM primary antibodies
  • Description

    Rabbit monoclonal [EPR23103-44] to SMARCA2 / BRM
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • WB: Wild-type HeLa, Daudi, HEK-293T, HeLa, MCF7 and Neuro-2a lysates. IHC-P: Human renal cell carcinoma, mouse cerebrum and rat kidney tissue. ICC/IF: HeLa and Neuro-2a cells. Flow Cyt (intra): HeLa cells.
  • General notes

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR23103-44
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromatin Remodeling
    • Swi / Snf
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Co-activators/co-repressors

Associated products

  • Alternative Versions

    • Anti-SMARCA2 / BRM antibody [EPR23103-44] - BSA and Azide free (ab269872)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human SMARCA2 (BRM) knockout HeLa cell line (ab265416)
  • KO cell lysates

    • Human SMARCA2 (BRM) knockout HeLa cell lysate (ab257687)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab240648 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
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 200 kDa (predicted molecular weight: 181 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/50.
Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 200 kDa (predicted molecular weight: 181 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/50.
Application notes
Is unsuitable for IP.

Target

  • Function

    Transcriptional coactivator cooperating with nuclear hormone receptors to potentiate transcriptional activation. 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. 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.
  • Involvement in disease

    Nicolaides-Baraitser syndrome
  • 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.
    Contains 1 QLQ domain.
  • Cellular localization

    Nucleus.
  • Target information above from: UniProt accession P51531 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6595 Human
    • Entrez Gene: 67155 Mouse
    • Entrez Gene: 361745 Rat
    • Omim: 600014 Human
    • SwissProt: P51531 Human
    • SwissProt: Q6DIC0 Mouse
    • Unigene: 298990 Human
    • Unigene: 644901 Human
    • Unigene: 491146 Mouse
    see all
  • Alternative names

    • ATP dependent helicase SMARCA2 antibody
    • ATP-dependent helicase SMARCA2 antibody
    • BAF190 antibody
    • BAF190B antibody
    • BRG1-associated factor 190B antibody
    • BRM antibody
    • FLJ36757 antibody
    • Global transcription activator homologous sequence antibody
    • hBRM antibody
    • hSNF2a antibody
    • MGC74511 antibody
    • Possible global transcription activator SNF2L2 antibody
    • Probable global transcription activator SNF2L2 antibody
    • Protein brahma homolog antibody
    • SMARCA2 antibody
    • SMCA2_HUMAN antibody
    • SNF2 alpha antibody
    • SNF2 like 2 antibody
    • SNF2-alpha antibody
    • SNF2/SWI2 like protein 2 antibody
    • SNF2L2 antibody
    • SNF2LA antibody
    • Sth1p antibody
    • Sucrose nonfermenting 2 like protein 2 antibody
    • SWI/SNF related matrix associated actin dependent regulator of chromatin subfamily A member 2 antibody
    • SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 2 antibody
    see all

Images

  • Western blot - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    Western blot - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    All lanes : Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : SMARCA2 knockout HeLa cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 181 kDa
    Observed band size: 200 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab240648 observed at 200 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab240648 was shown to react with SMARCA2 / BRM in wild-type HeLa cells in Western blot with loss of signal observed in SMARCA2 knockout cell line ab265416 (SMARCA2 knockout cell lysate ab257687). Wild-type HeLa and SMARCA2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab240648 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    Western blot - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    All lanes : Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648) at 1/1000 dilution

    Lane 1 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 181 kDa
    Observed band size: 200 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1: 3 minutes; Lanes 2-3: 26 seconds; Lane 4: 3 minutes.

    Two isoforms of SMARCA2/BRM are reported in human and mouse species (PMID:21811517).

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling SMARCA2/BRM with ab240648 at 1/2000 dilution (0.23 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat kidney. The section was incubated with ab240648 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)

    Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling SMARCA2/BRM with ab240648 at 1/2000 dilution (0.23 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse cerebrum. The section was incubated with ab240648 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)

    Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma tissue labeling SMARCA2/BRM with ab240648 at 1/2000 dilution (0.23 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human renal cell carcinoma. The section was incubated with ab240648 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

  • Flow Cytometry (Intracellular) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    Flow Cytometry (Intracellular) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling SMARCA2/BRM with ab240648 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labeling SMARCA2/BRM with ab240648 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing strong nuclear and weak cytoplasmic staining in Neuro-2a cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling SMARCA2/BRM with ab240648 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing strong nuclear and weak cytoplasmic staining in HeLa cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)
    Anti-SMARCA2 / BRM antibody [EPR23103-44] (ab240648)

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

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (0)

Publishing research using ab240648? Please let us know so that we can cite the reference in this datasheet.

ab240648 has not yet been referenced specifically in any publications.

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