All lanes : NONO Mouse mAb at 1/2000 dilution Lane 1 : THP1 cells membrane fraction Lane 2 : Jurkat cells membrane fraction Lysates/proteins at 20 ug per lane. Secondary Goat Anti-Mouse IgG-HRP, 5% milk conjugated at 1/10000 dilution Predicted band size : 54 KDa Observed band size : 54 KDa Blocking/Dilution buffer : 1x TBST.
NONO was immunoprecipitated from 1mg of THP1 cells membrane fraction, blotted with M25228 of 10 ug. Western blot was performed from the immunoprecipitate using M25228 at 1/1000 dilution. Anti-Mouse-IgG(HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/10000 dilution. Lane 1: THP1 cells membrane fraction. Lane2: IP product of THP1 cells membrane fraction.
Blocking and dilution buffer and concentration:5% milk/TBST.
Flow cytometric analysis of 2% paraformaldehyde-fixed THP1 (Human acute monocytic leukemia cell line) cells labeling NONO with M25228 at 1/200 dilution (red) compared with a mouse monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody, blue). Goat anti-mouse IgG (FITC) at 1/300 dilution was used as the secondary antibody.
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