Anti-SMARCA2 / BRM antibody - ChIP Grade (ab15597)

Overview

  • Product nameAnti-SMARCA2 / BRM antibody - ChIP Grade
    See all SMARCA2 / BRM primary antibodies
  • Description
    Rabbit polyclonal to SMARCA2 / BRM - ChIP Grade
  • SpecificitySpecific for human Brm from human cell lines (it does not appear to cross react with Brg1 which has been the nemesis of some other so-called Brm-specific antibodies).
  • Tested applicationsSuitable for: IP, ICC/IF, IHC-FoFr, ChIP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Mouse BRM/ SMARCA2 fusion protein derived from within residues 48-214 of the corresponding human sequence.(Note: Please see Muchardt C et al. EMBO J 15:3394-402 (1996) for further details).

  • Positive control
    • This antibody gave a positive signal in Mouse brain tissue lysate as well as HeLa and HEK293 whole cell lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab15597 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
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration. PubMed: 23359715
ChIP Use at an assay dependent concentration.
WB Use a concentration of 0.95 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 180 kDa).

Target

  • FunctionTranscriptional 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 diseaseNicolaides-Baraitser syndrome
  • Sequence similaritiesBelongs 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 localizationNucleus.
  • Information by UniProt
  • Database links
  • 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

Anti-SMARCA2 / BRM antibody - ChIP Grade images



  • Predicted band size : 180 kDa

    This image is courtesy of Gary Rosson

    The WB image shows a composite of 2 western blots illustrating the specificity of the ab15597. The cell lines used here are as follows:

     

    SW13- a human adrenal adenocarcinoma deficient in Brg1 and Brm (Neg.

    Control)

  • ab15597 staining BRM (red) in Rat E17 cortical neurons 7DIV by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.5% Triton + 3% BSA and blocked with 3% BSA for 30 minutes at 20°C. Samples were incubated with primary antibody (1/100 in PBS + 3% BSA) for 1 hour at 20°C. An Alexa Fluor® 555-conjugated Donkey anti-rabbit polyclonal (1/300) was used as the secondary antibody. Dapi is stained blue and MAP2 (1/500) stained green 

    See Abreview

  • A) ab15597 specifically immunoprecipitates Brm. Immunoprecipitations were performed on bacculovirus–expressed Flag-tagged Brm and Brg1 from insect cells.  2µg of ab15597 (left hand side) and 2µg of J1 (right hand side) were used for each IP. J1 is a polyclonal antibody recognising both, Brg1 and Brm.

    B) ab15597 chromatin immunoprecipitates Brm. Two cell lines 1 and 2 were treated with different stimuli and subjected to the ChIP procedure. Cells were fixed with formaldehyde for 10mins. The ChIP was performed with 2µg of 15597 or J1 and 10µl of protein G Dynal beads. The immunoprecipitated DNA was quanified by real-time PCR with primers specific for genes A and B, respectively. J1 is a polyclonal antibody recognising both, Brg1 and Brm.

  • Immunofluorescence using the Brm antibody ab15597.

    It appears the antibody works well in IF detecting BRM in D98oR WT cells. The D98oR BB10 blank omitted the primary antibody as a control. However, it was also positive on a cell line that has Brm knocked down by RNAi, (the BB10 cell line). This may be because RNAi is simply a knock down not a knock out or could be that the antibody is so good it is picking up the small amount of Brm remaining in the BB10s (highly possible), or the dilution is too high. Or, it may be that its crossreacting with another protein (though there was no cross rectivity with Brg1 in WB).

    Original magnification 400x. ab15597 dilution 1:100.

    Work is continuing on characterising this ab in IF.

  • All lanes : Anti-SMARCA2 / BRM antibody - ChIP Grade (ab15597) at 1 µg/ml

    Lane 1 : D98oR (subclone HeLa) positive control lysate
    Lane 2 : SW13 (Human adrenal adenocarcinoma) negative control lysate

    Lysates/proteins at 20 µg per lane.


    Predicted band size : 180 kDa
    Observed band size : 180 kDa
  • All lanes : Anti-SMARCA2 / BRM antibody - ChIP Grade (ab15597) at 1 µg/ml

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 180 kDa
    Observed band size : 220 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 35 kDa,60 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 5 minutes

References for Anti-SMARCA2 / BRM antibody - ChIP Grade (ab15597)

This product has been referenced in:
  • Takeshima H  et al. Frequent involvement of chromatin remodeler alterations in gastric field cancerization. Cancer Lett 357:328-38 (2015). Read more (PubMed: 25462860) »
  • Ceballos-Chávez M  et al. The chromatin Remodeler CHD8 is required for activation of progesterone receptor-dependent enhancers. PLoS Genet 11:e1005174 (2015). WB ; Human . Read more (PubMed: 25894978) »

See all 25 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Pancreas)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citric acid buffer
Permeabilization No
Specification Pancreas
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 27 2015

Thanks for your enquiry.

To help evaluate your current staining situation, I think it would be helpful to get some more complete details regarding your experimental procedures, sample preparation and IHC protocol.

Thanks in advance ...

Read More

We unfortunately were not able to obtain the identity of the cells and controls used by the collaborator who supplied the ChIP data.

Our own lab had this to say, based on the note below the ChIP data (...Immunoprecipitations were performed o...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa cells)
Loading amount 20 µg
Specification HeLa cells
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 40 minute(s) · Concentration: 4% · Temperature: 4°C
Username

Dr. Efrat Shema

Verified customer

Submitted Apr 04 2012

Thank you for providing more details. I have sent you a replacement antibody for ab15597 from a different lot as you requested. I hope that this new vial will work as stated for you. Your new order number is ********. Please let me know if there is any...

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Thank you for contacting Abcam. I am sorry that you were having issue with ab15597. I was just wandering what the actual problems were in WB, IHC and ChIP. Were you seeing no signal of high background? I will be very happy to send you a new aliquot of ...

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Thank you for contacting us. As discussed on phone we unfortunately do not sell small sizes of our products. The reason is that; we have over 70,000 products in our catalogue and having the small sized sample of each would be difficult to keep. How...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Rat E17 cortical neurons 7DIV)
Specification Rat E17 cortical neurons 7DIV
Fixative Formaldehyde
Permeabilization Yes - 0.5% triton & 3% BSA for 30 minutes
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Nov 22 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (MCF7 breast epithelial adenocarcinoma)
Specification MCF7 breast epithelial adenocarcinoma
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Detection step Real-time PCR
Positive control Region upstream of the transcription start site of the TFF1 gene
Region upstream of the transcription start site of the TK1 gene
Negative control Region on chromosome 12 (Untr12) that is far from any known gene annotation and not expected to be bound by BRM1
Username

Mr. Genpathway Inc

Verified customer

Submitted May 12 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Brain)
Loading amount 40 µg
Specification Brain
Gel Running Conditions Reduced Denaturing (8%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Xi Wang

Verified customer

Submitted Sep 10 2008

1-10 of 14 Abreviews or Q&A

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