Anti-SMARCA2 / BRM antibody (ab15597)
Key features and details
- Rabbit polyclonal to SMARCA2 / BRM
- Suitable for: ICC/IF, ChIP, WB
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-SMARCA2 / BRM antibody
See all SMARCA2 / BRM primary antibodies -
Description
Rabbit polyclonal to SMARCA2 / BRM -
Host species
Rabbit -
Specificity
Specific 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 Applications & Species
See all applications and species dataApplication Species WB MouseHuman
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Immunogen
Fusion protein corresponding to Mouse SMARCA2/ BRM aa 1-250.
(Peptide available asab171897) -
General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-SMARCA2 / BRM antibody (ab15597) 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)
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ChIP - Anti-SMARCA2 / BRM antibody (ab15597) This image is courtesy of Dr. Matthias Kaeser, RBIO-E/Emerson Lab, La JollaA) 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.
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Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody (ab15597) This image is courtesy of an anonymous abreview.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
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Immunocytochemistry/ Immunofluorescence - Anti-SMARCA2 / BRM antibody (ab15597) This image is courtesy of Garry Rosson, North Carolina at Chapel HillImmunofluorescence 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.
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Western blot - Anti-SMARCA2 / BRM antibody (ab15597)All lanes : Anti-SMARCA2 / BRM antibody (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
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Western blot - Anti-SMARCA2 / BRM antibody (ab15597)All lanes : Anti-SMARCA2 / BRM antibody (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
All lanes : 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
