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Our Abpromise guarantee covers the use of ab16088 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab16088 at 10ug/ml detecting APRIL by FACS analysis.
Left hand panel mock transfected cells
Right hand panel human APRIL tranfected cells
HEK 293 cells were mock transfected or transfected with an expression plasmid enabling surface expression of a non-cleavable human APRIL. Cells (5x105) were incubated on ice for 30 min in 50 ul FACS buffer (PBS, 5% Fetal calf serum, 0.02% azide) containing 10ug/ml of Aprily-1 antibody. After washing in FACS buffer, FITC-conjugated antibody to mouse IgG was added. Cells were incubated on ice for 30 min., washed and analyzed by flow cytometry.
10ug of protein was applied to the gel. Revealed with ab16088 (1ug/ml) and HRP-coupled anti-mouse secondary antibody (1:4000).
The human APRIL construct used in this experiment is an uncleavable fusion protein between human BAFF (aa 1-132) and human APRIL (aa 93-233).