Proteomics Strategy for Identifying Candidate Bioactive Proteins in Complex Mixtures: Application to the Platelet Releasate
Figure 3
Fractionation of the platelet releasate by ion-exchange chromatography. (a) From each releasate fraction collected, 80 L was precipitated to remove salt, resuspended in 10 L of Laemmli buffer, and separated by MW on 4%–20% acrylamide gels. Silver-stained gels are displayed below corresponding sections of the ion-exchange UV detected chromatogram. A UV wavelength of 280 nm was used to detect eluting peptides. (b) The number of spectra detected in each fraction following LC MS/MS of tryptically digested 1D gel pieces. (c) The number of proteins identified in each fraction using SEQUEST with a score greater than 0.9. (d) Representation of the number and proportion of proteins in a single fraction, two adjacent fractions; two nonadjacent fractions, or more than two fractions.
