SUPPORTING INFORMATION A Universal Flow Cytometry Assay for Screening Carbohydrate-Active Enzymes Using Glycan Microspheres Aarthi Chandrasekaran, *,a,b Kai Deng, a,b Chung-Yan Koh, b Taichi Takasuka, c Lai F. Bergeman, c Brian G. Fox, c Paul D. Adams, a,d Anup K. Singh* ,a,b a Technology Division, Joint BioEnergy Institute, Emeryville, CA; b Biotechnology and Bioengineering Department, Sandia National Laboratories, Livermore, CA; c Great Lakes Bioenergy Research Center, University of Wisconsin, Madison, WI; d Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA. * Corresponding Author Emails: achandrasekaran@lbl.gov ; aksingh@sandia.gov Table of Contents Supporting Data………………………………………………………………………………...S2 Figure S1: Process flow chart for glycan microsphere-based assay Figure S2: Schematic of enzymatic activities of GTs: GalT and 6-ST Figure S3: Detection sensitivity of 6-ST assay Figure S4: Validation of covalent conjugation of glycan substrates to microspheres Figure S5: Schematic of cellulase activity Materials and Methods…………………………………………………………………………S7 Materials and Reagents Recombinant Cellulase Expression Glycan Substrate Synthesis Figure S6: The structures of four glycan-C5-NH 2 substrates used in this study Figure S7: Synthesis of building blocks compounds 7 and 9 Figure S8: Synthesis of azide modified cellotetraose compound 3 Figure S9: Synthesis of azide modified cellobiose compound 4 Glycan Substrate Conjugation to Microspheres Enzyme Assays and Detection References……………………………………………………………………………………..S21 Proton NMR Spectra of Glycan Compounds……………………………………………….S22 Electronic Supplementary Material (ESI) for Chemical Communications This journal is © The Royal Society of Chemistry 2013