Quantitation of phosphopeptides binding to and recovered from different types of IMAC material.32P-labeled phosphopeptides were produced by in vitro autophosphorylation.

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Quantitation of phosphopeptides binding to and recovered from different types of IMAC material.32P-labeled phosphopeptides were produced by in vitro autophosphorylation of Arabidopsis microsomes and purified with the indicated IMAC materials. Quantitation of phosphopeptides binding to and recovered from different types of IMAC material.32P-labeled phosphopeptides were produced by in vitro autophosphorylation of Arabidopsis microsomes and purified with the indicated IMAC materials. Phosphate buffer or dilute ammonia with different pH values, with and without addition of 30% acetonitrile, was used for elution. Phosphopeptides were quantified by spotting onto small filter papers and PhosphorImage analysis. “Bound” corresponds to input minus flowthrough. The bars for IDA/NTA indicate unspecific binding to the chelating resins without bound metals. Thomas S. Nühse et al. Mol Cell Proteomics 2003;2:1234-1243 © 2003 The American Society for Biochemistry and Molecular Biology