Instructions for Use - Glyco Kit MB-LAC Con A - Bruker

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Isolation of glycopetides from tryptic RNase B digest Sample: Tryptic digest of RNase B (10 pmol) Beads: MB-LAC Con A (10 µl) Buffers: BB 2, WP 2.1, WP 2, EB Target: MTP AnchorChip 600/384 Matrix: 2,5-DHB MALDI mode: Reflector 8000 2224.014 2760.078 Total digest Intens. [a.u.] 6000 4000 2000 0 1200 1070.410 1150.567 1287.584 1399.618 1504.603 1691.612 1934.747 2123.800 2170.808 2286.860 2364.870 2421.885 2517.165 H H H H 2672.212 2712.085 2817.102 2896.080 3023.353 3241.313 3332.334 Eluate 1000 800 1934.837 2420.982 600 400 200 SRNLTK 1991.847 2096.882 2258.926 2364.905 2477.985 2583.029 0 1250 1500 1750 2000 2250 2500 2750 3000 3250 3500 m/z The glycosylated peptide RNase B aa58-63 was successfully isolated from the complex total digest of RNase B. The peak at 1934.8 Da correlates to the depictured glyco-peptide structure comprising five mannose residues. Four additional signals each with a distance of 162 Da (one hexose unit) to the previous peak correspond to higher mannosylated forms of the same peptide comprising six, seven, eight or nine mannose residues, respectively. Page 18 of 21
References 1 Sparbier K, Koch S, Kessler I, Wenzel T, and Kostrzewa M.(2005). Selective Isolation of Glycoproteins and Glycopeptides for MALDITOFMS Detection Supported by Magnetic Particles. J Biomol Tech, 16: 407 – 413. 2 Sparbier K, Wenzel T, Kostrzewa M. (2006). Exploring the binding profiles of ConA, boronic acid and WGA by MALDI-TOF/TOF MS and magnetic particles. J ChromB, 840: 29–36. 3 Sparbier K, Asperger A, Resemann A, Kessler I, Koch S, Wenzel T, Stein G, Vorwerg L, Suckau D, Kostrzewa M. (2007). Analysis of Glycoproteins in Human Serum by Means of Glyco-specific Magnetic Bead Separation and LC-MALDI-TOF/TOF Analysis with Automated Glycopeptide Detection. J Biomol Tech, Sept., accepted for publication. 4 Sparbier K; Kessler I, Koch S, Mix G, Kostrzewa M. (2007). CLINPROT Glyco-beads: Verification of Long-Term Stability and Batch-to-Batch Reproducibility. Technote Bruker Daltonic (tn-20). 5 Drake RR, Schwegler EE, Malik G, Diaz J, Block T, Mehta A, and Semmes OJ. (2006). Lectin Capture Strategies Combined with Mass Spectrometry for the Discovery of Serum Glycoprotein Biomarkers. Mol&CellProt, 5.10:1957 – 1967. 6 Kamra A, GuptaMN. (1987). Crosslinked concanavalin A-O-(diethylaminoethyl)-cellulose--an affinity medium for concanavalin A-interacting glycoproteins. Anal. Biochem., 164: 405 - 410. 7 Yahara I, Edelman GM. (1972). Restriction of the mobility of lymphocyte immunoglobulin receptors by concanavalin A. PNAS, 69: 608 - 612. 8 Goldstein IJ, Hollerman CE, SmithEE. (1965). Protein-Carbohydrate Interaction. II. Inhibition Studies on the Interaction of Concanavalin A with Polysaccharides. Biochemistry, 41: 876 -883. 9 Baumann S, Ceglarek U, Fiedler GM, Lembcke J, Leichtle A, Thiery J. (2005): Standardized approach to proteome profiling of human serum based on magnetic bead separation and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. Clin Chem., 51, 973-80. 10 Fiedler GM, Baumann S, Leichtle A, Oltmann A, Kase J, Thiery J, Ceglarek U. (2007): Standardized peptidome profiling of human urine by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Clin Chem. 53, 421-8. Page 19 of 21
Page 1 and 2: CARE products are designed for supp
Page 3 and 4: Table of Contents General Informati
Page 5 and 6: Kit Components Materials provided i
Page 7 and 8: Recommended sample preparation proc
Page 9 and 10: 14. Transfer the tube from the Magn
Page 11 and 12: MALDI-TOF MS Target Preparation MAL
Page 13 and 14: Matrix preparation on the MALDI-MS
Page 15 and 16: 7. After drying of the spots the ta
Page 17: Examples Isolation of a small glyco
Page 21: Ordering Information Product Size (

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