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. Author manuscript; available in PMC: 2010 Jul 13.
Published in final edited form as: Mol Cell Proteomics. 2007 Jan 5;6(4):669–676. doi: 10.1074/mcp.M600218-MCP200

Fig. 1. Optimizing the batchwise IMAC protocol using 32P-labeled peptides.

Fig. 1

A, effect of solvent on phosphopeptide solubility. B, combined loading and washing loss at different loading pH values. C, combined loading and washing loss at different organic solvent concentrations in washing solutions. D, elution efficiency of phosphopeptides from IMAC beads with buffers containing 50 mm Na2HPO4, 50 mm EDTA, or 5% TFA. E, elution efficiency of phosphopeptides from IMAC beads with buffers containing organic solvent. Experimental procedures for the experiments are described in detail in the supplemental information. HAc, acetic acid; MeOH, methanol.