Sigma-Aldrich
Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) antibody produced in mouse
A8592
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A FLAG-tagged construct was expressed in HEK293T cells and 50 µg of clarified lysate was loaded for SDS-PAGe followed by Western Blotting with a FLAG-HRP conjugated antibody. The goal was to test this antibody so that one can exclude secondary HRP conjugated antibody step and save one hr of incubation.
Western Blot
HEK293T cells
1:2000 dilution in blocking buffer for 2 hr at room temp
5% Milk powder in 1xTBS-T (0.1% Tween)
NA
ECL for 5 sec on a film
FLAG-tagged Kif7 was expressed in HEK293T cells and could be nicely detected by a HRP-conjugated FLAG antibody
None
Eliminates secondary antibody step
Nice antibody that saves 1 hr of secondary antibody step and 15-30 min wash steps.