NEW PRODUCTS N-Glycosidase F Deglycosylation Kit addition, the action of oligosaccharide pro- and Endoglycosidase H cessing inhibitors, such as castanosper- Deglycosylation Kit mine- and deoxymannojirimycin, on a gly- Fast and easy deglycosylation of glycan coprotein can be followed. chains on glycoproteins For structural analysis of N-(aspar- The N-Glycosidase F Deglycosylation agine)-linked carbohydrate chains of Kit can be used to test for the existence glycoproteins, chemical hydrazinolysis of asparagine-linked glycan chains on and enzymatic methods are most widely glycoproteins. The kit enables a researcher employed to cleave all common classes of to estimate the number of glycan chains oligosaccharides. During hydrazinolysis, Figure 15 Reaction principles of (A) Endoglyco- bound to a protein. After the deglycosy- however, the protein is destroyed, and sidase H Deglycosylation Kit and (B) N-Glyco- sidase F Deglycosylation Kit. lation reaction, the protein can be degradation and modification of the analyzed by SDS-PAGE (Figure 14); a released sugar chains have been observed. Both new kits are time saving (only shift to a lower apparent molecular Enzymatic procedures are, therefore, the 1 hour incubation time) and convenient weight after the reaction indicates the only methods allowing the structural (only 3 easy to perform working steps; all existence of asparagine-linked glycan analysis of the glycan and the protein part. necessary buffers and controls are chains. The glycan, as well as the protein, Several endoglycosidases useful for these included). Unlike with chemical proce- moiety can also be used for further struc- applications have been described; these dures, both the glycan part and the protein tural and functional analysis. enzymes are useful tools for the selective part can be analyzed after the reaction. The new Endoglycosidase H Deglyco- removal and characterization of the indi- Each kit has proven its reliability in a func- sylation Kit is useful for testing the exis- vidual glycan types. Endoglycosidase H tion test. tence of “high mannose” type and (Figure 15) and endoglycosidase F1 (not Product Cat. No. Pack Size “hybrid” type asparagine-linked glycan included in the kit) act only on “high man- N-Glycosidase F 1 836 552 1 kit for chains on purified glycoproteins. Like nose” type and “hybrid” type structures, Deglycosylation Kit 12 reactions the N-Glycosidase F Deglycosylation Kit, while endoglycosidase F2 (not included in Endoglycosidase H 1 836 579 1 kit for it allows the estimation of the number of the kit) prefers “biantennary complex” Deglycosylation Kit 20 reactions glycoprotein-bound glycan chains via type chains. N-glycosidase F is able to deglycosylation reaction and SDS-PAGE release all common classes of N-glycans Also Available Cat. No. Pack Size (Figure 14). It serves also as a useful tool from the protein backbone; the enzyme is Endoglycosidase H 1 088 726 200 mU (200 µl) for the study of glycoproteins during their not a glycosidase but rather an amidase, so 1 643 053 500 mU (500 µl) synthesis, processing, and secretion. In it converts asparagine to aspartic acid. Endoglycosidase F1 1 636 197 3 U (60 µl) Endoglycosidase F2 1 694 413 20 mU ( 50 µl) 123 123 O-Glycosidase 1 012 142 25 mU 1 012 169 100 mU N-Glycosidase A 1 642 995 5 mU (0.1 ml) Trans. Carb. N-Glycosidase F 1 365 185 20 units 1 365 193 50 units 1 643 045 250 units Enz. Enz. DIG Glycan 1 142 372 1 kit Detection Kit DIG Glycan 1 210 238 1 kit ac.gp. Differentiation Kit DIG Glycan ELISA 1 460 366 1 kit RNase RNase DIG Glycan/Protein 1 500 783 1 kit Double Labeling Kit Figure 14 Control glycoproteins digested with (A) N-Glycosidase F Deglycosylation Kit; Lane 1: digested gly- coproteins; Lane 2: undigested glycoproteins; Lane 3: protein molecular weight marker. (B) Endoglycosidase H Deglycosylation Kit; Lane 1: protein molecular weight marker; Lane 2: undigested glyco- proteins; Lane 3: digested glycoproteins. BIOCHEMICA ■ NO. 4 [1996] 13 CONTENTS.