Method for in-situ desalination and enrichment on trace amount of protein or polypeptide target
A protein and trace technology, applied in the field of biochemical analysis, can solve the problems of insufficient elution, affecting detection sensitivity, and not suitable for large-scale applications of proteomics, achieving high sensitivity, high reproducibility, and improved sensitivity Effect
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[0028] After the protein from rat liver was separated by 2D-gel and stained with Coomassie Brilliant Blue, 10 points were selected, and the labels of the points were represented by 1 to 10 according to the staining depth from deep to light ( Figure 5 ), followed by enzymolysis with trypsin. First, the disc on the MALDI target plate is coated with a block polymer of microscopic phase separation structure. After the polymer dot film is formed, 0.5 microliters of enzymatic peptide sample solution (solvent is 50% acetonitrile, 50% Water, 0.1% trifluoroacetic acid) were spotted on the coating dot, another 0.5 microliter of the same enzymatic peptide sample solution was dotted on the light target dot, and dried naturally at room temperature; finally, the volume of 0.5 microliter The organic matrix solution (2-cyano-4-hydroxycinnamic acid (α-CHCA), the solvent is 50% acetonitrile, 50% water, 0.1% trifluoroacetic acid) was spotted on the surface of the sample, and dried naturally at ...
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