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Synthetic peptide conjugated to KLH derived from within residues 1400 - 1500 of Human BRG1.
(Peptide available as ab13736.)
Our Abpromise guarantee covers the use of ab4081 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent dilution. PubMed: 20522713|
|WB||1/500 - 1/1000. Detects a band of approximately 235 kDa (predicted molecular weight: 185 kDa).|
Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: BRG1 knockout HAP1 cell lysate (40 µg)
Lane 3: HeLa cell lysate (40 µg)
Lane 4: K562 cell lysate (40 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab4081 observed at 240 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab4081 was shown to recognize BRG1 when BRG1 knockout samples were used, along with additional cross-reactive bands. Wild-type and BRG1 knockout samples were subjected to SDS-PAGE. Ab4081 and ab18058 (loading control to Vinculin) were diluted at 1/500 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) with Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.
ab4081 staining Brg1 in U2OS cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with NP-40 and blocked with 3% BSA for 1 hour at 21°C. Samples were incubated with primary antibody (1/200 in PBS + 3% BSA) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG polyclonal was used as the secondary antibody (1/500).
Rabbit polyclonal to BRG1 (ab4081) at 1/500 on HeLa Nuclear extract (20 ug per lane).
Lane 1: ab4081 (1/500)
Lane 2: ab4081 (1/500) + blocking peptide (1 ug)
Secondary antibody: Goat anti-rabbit (HRP) - ab6721
This image is courtesy of an anonymous abreview.Whole cell lysate prepared from HeLa cells was loaded at 150000 cells.ab4081 used at a 1/1000 dilution.The secondary used was an HRP conjugated donkey polyclonal used at a 1/5000 dilution.
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