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PƼA&SJ8,AL Ā[}k`0ho:5\c e(hX2_)r9 &بb 4,h(y }4S<'xm' 3p@|ޅOzMdMw퓛7tT`$Ѕ 7?rtn|-=0/N11~fЎ~nh1ޏ6G>8Z(p;un_ ,3N3_);ohFs t_ /?S~I[wo~}(W^h5j0J?<̨sYi S O m#y>/~6ЍݼZCᏭtp^}+|ewy hxGmnhlDF+2Tb9_|ˀ|~4ֳ 1<)ď'aplFӂ<' ?~,,Phosphopeptide identification Beta casein was used to examine the capacity of Phos-Pep TM , non-IMAC phosphopeptide enrichment kit. Beta casein (purchased from Sigma, Cat.No.C6905) was digested with trypsin and phosphopeptide was enriched using Phos-Pep TM and peptide map was measured. Phosphopeptide which was not shown in total digest of beta casein(Fig.2.) was detected in enriched fraction(Fig.2. lower panel, Fig.3. A). Among these phosphopeptide, mono phosphopeptide(2062) and tetra- phosphopeptide(3124) was originated from beta casein. Some phosphopeptide, 1660, and 1952, seems to be originated from contaminated alpha casein, because this phosphopeptide is a major component of commercially available alpha casein phosphopeptide(see Fig.4.). Some peptides, ranging 2884 to 3054, seems to be the derivatives of tetra-phophopeptide of 3124, which have differential mass according to the status of the number of phosphorylation site. Some of phosphopeptide was identified by PDS of MALDI-TOF by detecting the mas loss of phosphoric or phosphorous acid (Fig.3.). Fig.3. Identification of phosphopeptide enriched from beta casein trypsin digest by PSD(post source decay) using MALDI-TOF. Panel A represents the MALDI-TOF spectrum of enriched phosphopeptide from beta casein trypsin digest. Blue asterisk represent the enriched phosphopeptides. Rest of the seven spectrums represent the PSD spectrum of enriched phosphopeptide. 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