Hello

I use Sigma's polyclonal Ab that detects Y1092 of pEGFR.

I don't think there's a problem with the Ab because it works in other cells.

My question is - maybe the problem is that there's no/not enough over-expression of EGFR in DLD-1 cells (I could not find the answer for that online)? And maybe this is the reason?

I detect fine the total amount of EGFR but when trying pEGFR the blot is blank.

I tried changing a lot of technical things but nothing seems to work.

Plus, adding EGF for 5 mins before harvesting helps, but the bands are wide and blurry and it interferes with a treatment's effect I want to check.