Proteomics Strategy for Identifying Candidate Bioactive Proteins in Complex Mixtures: Application to the Platelet Releasate
Figure 2
A pro-migratory effect identified in unfractionated and fractionated platelet releasate. THP-1 monocyte cell migration was measured using a Boyden chamber assay over 2 hours. (a) Assessment of migrated cells adhering to the underside of the membrane in response to platelet releasate, Serum (10%), and MCP-1 (100 ng/mL). = 3; (b) Adherent migrated THP-1 cells in response to 10, 25 or 100 L platelet releasate; = 3. (c) Fractions were pooled in groups of three and added to serum-free media (SFM) in the bottom chamber. Positive controls include MCP-1 and unfractionated platelet releasate; = 3. (d) Localisation of the pro-migratory effect of Fractions 25–27 to an individual fraction. Each fraction (2 L) was added to serum-free media in the bottom chamber of a separate well and compared with the combination of Fractions 25–27 (2 L of each fraction). TRAP = thrombin receptor-activating peptide. = 3. The average number of migrated cells adhering to the underside of the membrane was calculated from ten 40X fields of view.
