Figure 5: Phosphorylation profiling of intracellular proteins by using an antibody microarray system. (a) Comparative data obtained by probing with HRP–SA only before (left, –) and after (right, +) adding a blocking treatment. (b) Profiling of multiplex protein phosphorylation involved in the LPS/PMA-signaling pathway of Raw 264.7 cells. Images from lysates before (control, –) and after (+) LPS/PMA treatment are shown in the left-hand and right-hand panels, respectively. (c) Densitometric data from the images shown in b. The reported values of signal intensity are means in duplicate. Proteins having more signal intensities with values in excess of 20,000 and which also increased by more than a factor of two after LPS/PMA treatment are shown. All the ECL images were obtained by using the Lumigen TMA-6 and LAS 3000 image analyzer, and the densitometric analysis was performed by using the Multi Gauge software attached to the analyzer. [Reprinted with permission from Ref. 29 ©(2013) Elsevier Inc.].