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文献和实验the protein sample.MALDI targetMicrocentrifugeMicrocentrifuge tubes (standard and 0.7-mL)METHODFigure 1. Schematic of affinity binding of phosphopeptides to immobilized metal ion affinity columns.Preparation of the Immobilized Metal Ion Affinity Column Insert
Characterization of Phosphopeptides Using a Combination of Immobilized Metal Ion
of the phosphopeptides from the metal ion column prior to MS analysis can result in sample loss. Affinity-bound analytes, including phosphopeptides, can be directly analyzed by MALDI-MS without prior elution from the affinity media. Also, consecutive enzymatic reactions
Affinity chromatography (with HIS-tagged proteins)
minutes on ice, wash the resin twice in 1mL of AC buffer to remove unbound protein and to equilibrate the resin in AC buffer. 12) Set up the columns. Add 50uL of settled affinity resin to the prepared tips. Allow the resin to settle on ice. 13
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