FACEÂ® N-Linked Oligosaccharide Profiling Kit - ProZyme

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FACE N-Linked Oligosaccharide Profiling Kit (80 Reactions) NOTE: We want successful results for our customers, so please read this entire booklet before starting your experiment. Contents Page No Kit Contents 2 Additional Reagents and Equipment Required 3 Introduction 4 Principle Description of FACE N-linked Profiling System Nature of FACE Oligo Standards and Controls Summary of Protocol Protocols Section 1: Enzymatic Release of Oligosaccharides from Glycoproteins 9 Section 2: Labeling of Oligosaccharides from Glycoproteins 11 Section 3: Preparation of Samples and Standards for Electrophoresis 12 Section 4: FACE Electrophoresis 14 I. Preparation of OLIGO Gel Running Buffer II. Set-up of FACE Electrophoresis Apparatus III. Loading and Running FACE N-Linked Profiling Gels Section 5: Processing of FACE N-Linked Profiling Gels 18 I. Gel Imaging using UV Transilluminator (lightbox) II. Gel Imaging using FACE Imaging System III. Gel Handling IV. Oligosaccharide Preparation Troubleshooting Guide 21 Figures 23 Page 1
Kit Contents Catalog # Code Component Quantity Storage GK60000 10 each Oligo Profiling Gels precast gels Room Temp WS0007 Oligo Gel Running Buffer 5 packs Room Temp on arrival GK50050 4 packs GK50004 2 packs 4°C (when reconstituted) E1 2x Loading Buffer 1 tube Room Temp E2 Tracking Dye 1 tube Room Temp C1 N-Glycanase® 1 tube (100 mU) 4°C C3 2x Profiling Enzyme Buffer 1 tube 4°C C11 2% SDS/1M β-mercaptoethanol 1 tube 4°C C12 15% NP-40 1 tube 4°C L1 Reducing Agent (NaBH 3 CN) 2 tubes Room Temp on arrival -70°C (when reconstituted) L2 Oligo Labeling Dye (ANTS) 2 tubes Room Temp on arrival -70°C (when reconstituted) L3 Oligo Labeling Diluent 2 tubes Room Temp L4 Labeling Solvent (DMSO) 2 tubes Room Temp GK50006 E3 Oligo Ladder Standard 1 tube -20°C E4 Oligo Profiling Control 1 tube -20°C E5 Oligo Quantitation Control 1 tube -20°C on arrival -70°C (when reconstituted) Page 2
Page 1 and 2: FACE ® N-Linked Oligosaccharide Pr
Page 6 and 7: Additional Reagents and Equipment R
Page 8 and 9: Description of the FACE N-linked OL
Page 10 and 11: OLIGO Quantitation Control (E5) The
Page 12 and 13: Protocols SECTION 1 ENZYMATIC RELEA
Page 14 and 15: SECTION 2 LABELING OF OLIGOSACCHARI
Page 16 and 17: 3a Load 4 µl directly onto the gel
Page 18 and 19: 2 Pour pre-cooled OLIGO Gel Running
Page 20 and 21: 14 When the electrophoresis is comp
Page 22 and 23: 2 Develop film according to manufac
Page 24 and 25: TROUBLESHOOTING GUIDE Sample not mo
Page 26 and 27: Separation of N-Linked Oligosacchar
Page 28 and 29: 1 2 Figure 4. Lane 1 E3 - FACE Oli

FACEÂ® N-Linked Oligosaccharide Profiling Kit - ProZyme

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