Figure 5

Exposure time and protein amount is important for accurate measurement of signal. The intensity of signal (AUC) for total and phosphorylated 4EBP1 (A) and total Akt 1/2/3 (B & C) was examined at exposure times between 30–960 seconds in MV4-11, MOLM-14, OCI-AML3 and HL60 cells. Higher exposure times (240–960 secs) resulted in signal burn-out which was not observed at lower exposure times (30–120 secs). More abundant proteins such as 4EBP1 exhibit improved linearity in signal with titration of protein loading (Additional file 1: Figure S1).