Figure 2
Comparison between lanthanum and micro-column PBMCs’ phosphoprotein profiles. (A) Sequential staining of PBMCs’ phosphoproteins enriched by LaCl3 (La) and micro-column (μ-clmn) and Western blot investigation of phosphoproteins using an anti-phospho-Ser/Thr/Tyr monoclonal antibody. Eluted proteins were separated by 1D-SDS-PAGE, and stained first with ProQ to visualize phosphoproteins, then with SYPRO to visualize total proteins. (B) The same analysis was performed on the corresponding flow-through. (C) Two identical aliquots of PBMCs protein lysate were both incubated with alkaline phosphatase (AP) in the same conditions. Then, the two aliquots were adjusted at pH = 7.4 (physiological pH) before being treated with lanthanum or micro-column. The entire amount of isolated proteins were separated by 1DE and stained first with ProQ, then with SYPRO. AP: alkaline phosphatase; PL: PBMCs protein lysate; PL + AP: dephosphorylated PL. MW markers: PeppermintStick™ Phosphoprotein MW Standards.