Enzyme-linked immunosorbant assays for glucosidases involved in processing of N-linked glycoproteins

Conference · Fri May 01 04:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6156936

Nakhasi, D; Saxena, E; Shailubhai, K; Alston-Mills, B; Vijay, I K

The conventional biological assays for glucosidase I and II are to determine the release of UC-glucose from radiolabeled Glc3Man9(GlcNAc)2 and Glc/sub 1-2/Man9(GlcNAc)2, respectively by the actions of glucosidases. They have developed ultrasensitive ELISA's for these two glycosidases by using highly specific polyclonal antibodies prepared against purified glucosidase I and II from the bovine mammary tissue. Anti-glucosidase I and anti-glucosidase II antibodies are shown to be monospecific and are cross-reactive towards glucosidase I and II from liver, mammary gland, kidney and heart tissues of cow, guinea pig, mouse and rat, respectively. Western blot analysis revealed that anti-glucosidase I and anti-glucosidase II are very specific and are not cross-reactive towards any other protein present in solubilized microsomal extracts. Using these antibodies, conditions for ELISA's for glucosidase I and II were standardized. Dose response curves for purified enzymes and crude extracts were developed. A correlation between biological activities of these enzymes and immunoreactive glucosidase I and II, present in the solubilized microsomal preparations of liver, mammary gland, kidney and heart tissues from cow, guinea pig, mouse and rat, is made. The sensitivity of ELISA's for glucosidase I and II are in the range of 10 - 1000 ng of immunoreactive species.

• Research Organization: Univ. of Maryland, College Park
• OSTI ID: 6156936
• Report Number(s): CONF-870644-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 46:6
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
ANIMALS
ANTIBODIES
BODY
CARBOHYDRATES
CARBON 14 COMPOUNDS
CARDIOVASCULAR SYSTEM
CATTLE
DIGESTIVE SYSTEM
DOMESTIC ANIMALS
DOSE-RESPONSE RELATIONSHIPS
ENZYMES
GLANDS
GLUCOPROTEINS
GLUCOSE
GLUCOSIDASE
GLYCOSYL HYDROLASES
GUINEA PIGS
HEART
HEXOSES
HYDROLASES
ISOTOPE APPLICATIONS
KIDNEYS
LABELLED COMPOUNDS
LIVER
MAMMALS
MAMMARY GLANDS
MICE
MONOSACCHARIDES
O-GLYCOSYL HYDROLASES
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RADIOENZYMATIC ASSAY
RATS
RODENTS
RUMINANTS
SACCHARIDES
TRACER TECHNIQUES
VERTEBRATES