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Merck KGaA
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10 二月 2012
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Protein glycosylation influences crucial biological and pharmaceutically relevant events. Glycoanalysis studies of antibodies, receptors and other glycosylated proteins require highly sensitive, selective, efficient and reproducible methods for sample preparation and mass spectrometry (MS). Effective analysis requires glycopeptide enrichment to compensate for low abundance glycopeptides in complex mixtures and, subsequently, to enable glycan and attached peptide analysis by MS.
The ProteoExtract® Glycopeptide Enrichment Kit is designed to capture high-mannose, hybrid, and complex type glycosylated peptides from complex protein mixtures. The kit uses a proprietary ZIC® Glycocapture Resin. In combination with the included optimized buffers, the ZIC Glycocapture Resin is highly selective for glycosylated peptides in the presence of abundant non-glycosylated peptides. The protocol is optimized to allow efficient removal of non-glycosylated peptides with an almost complete glycopeptide recovery (as quantitatively assessed by N-Glycan mapping). It allows both site-specific determination of the glycan structure and parallel analysis of the attached peptides. Glycopeptide enrichment is non-specific and nonselective, without preference for particular glycan structures and without loss of distinct glycans. The enriched glycopeptide fraction is directly compatible with downstream analysis by MALDI and ESI-MS. One ProteoExtract® Glycopeptide Enrichment Kit provides sufficient reagents for 50 enrichment reactions. 10 µl trypsin-digested protein sample with maximum glycopeptides concentration 2-4 µg/µl can be processed in one reaction.
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| Storage | +2°C to +8°C | ||
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Ambient or Blue Ice
Flammable |
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| 数据 | |||
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 The Glycopeptide Enrichment Kit allows for selective and sensitive glycopeptide enrichment with low background from complex mixtures. A complex peptide mixture derived from a tryptic digest of bovine fetuin, α1 acid glycoprotein, RNaseB, histone, BSA, and human IgG was processed using the ProteoExtract® Glycopeptide Enrichment Kit as outlined in the User Protocol TB527. Mass spectrometry analysis was performed using ESI-LC/MS equipment operated in positive mod. A. Sum spectra of unprocessed sample. B. Sum spectra of PNGaseF digested glycopeptides recovered using the ProteoExtract Glycopeptide Enrichment Kit. |
