Enzyme-linked immunosorbent assay (ELISA)-based quantification and identification of in vitro enzyme-catalyzed glycosphingolipid synthesis and degradation products with carbohydrate sequence-specific monoclonal antibodies
✍ Scribed by Cheryl L.M. Stults; Bruce J. Wilbur; Bruce A. Macher
- Publisher
- Elsevier Science
- Year
- 1988
- Tongue
- English
- Weight
- 531 KB
- Volume
- 174
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A new method has been developed to monitor glycosyltransferase and glycosylhydrolase activities. Reaction product identification and quantification are accomplished simultaneously with an enzyme-linked immunosorbent assay (ELISA) using carbohydrate sequence-specific monoclonal antibodies. P-Galactosyltransferase and cu-galactosidase reactions were used to illustrate the salient features of the method. These include simple product identification and quantitication, no detergent requirement, consumption of small amounts of reagents, and no use of radioisotopes. Furthermore, it is possible to measure substrate disappearance or product formation with this method. Enzyme characteristics such as K,, V,,,,, divalent cation requirement, and pH optimum were investigated with this new method. 0 1988 Academic press, hc.