Post Wed Nov 26, 2014 9:04 am

Problem with pErk staining and signal intensity?

FDCP1 cells were stimulated with IL3 and stained for pErk Yb171, pStat5 5Nd150 and pAkt Sm152.
The basal level of pErk appears to be not stained properly and very less cells are possitive to pErk and signal for stimulated pErk is too high. I am sure that i have added same amount of antibody and same amount of cells in both tubes. here is the screen shots. I am thinking what would be the reason ? .
Screen Shot 2014-11-26 at 9.58.51 AM.png
contour map
Screen Shot 2014-11-26 at 9.56.56 AM.png
Histogram overlay