geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

A simple and specific assay of glycosyltransferase and glycosidase activities by an elisa method and its application to assay of galactosyltransferase activity in sera from patients with cancer


, : A simple and specific assay of glycosyltransferase and glycosidase activities by an elisa method and its application to assay of galactosyltransferase activity in sera from patients with cancer. Journal of Biochemistry (Tokyo) 107(3): 493-498

A simple, sensitive, and specific assay method for glycosyltransferase and glycosidase activities has been established by means of an enzyme-linked immunosorbent assay (ELISA) using monoclonal antibody, H-11 directed to lactoneotetraosylceramide (nLC4Cer). Enzyme activity was determined by assaying the amount of reaction product, nLC4Cer with the ELISA method. For the assay of galactosyltransferase activity, lactotriaosylceramide (Lc3Cer) immobilized on a 96-well microtiter plate was incubated with bovine milk galactosyltransferase in cacodylate buffer (pH 6.8) containing Triton CF-54, Mn2+, and UDP-galactose. Optimum incubation conditions for the enzyme were determined. Glycosidase activity was also assayed by the ELISA method by using Clostridium perfringens sialidase and neolacto-series gangliosides as substrates, and the substrate specificities towards the gangliosides were examined. By this method, 3-100 pmol of reaction product could be determined. The assay method has several advantages as follows: 1, the method is simple; 2, separation of the reaction product is not required; 3, quantification and identification of the reaction product were done simultaneously; 4, naturally occurring substrates are available (especially for glycosidase); 5, many samples can be assayed in one microplate; 6, sensitivity is very high. The present method was applied for the detection of galactosyltransferase in human sera. Significant elevations of the galactosyltransferase levels were observed in the sera from cancer patients. The formation of nLC4Cer was confirmed by employing the TLC-immunostaining method for bands of Lc3Cer after incubation of the bands with serum and cofactors on an HPTLC plate.

US$29.90


Other references

Angelova-Gateva, P.; Kalinkova, G., 1978: Infrared spectroscopy of erythrocyte membranes. The authors examined the properties of erythrocyte membranes, obtained from young and old erythrocytes of 2-old rats by means of infrared spectroscopy. The films, prepared on CeF2 plates were recorded before and after 30 minutes treatment with met...

Jones, L., 1996: Ethics case study: double exposure. Probe 30(3): 111-111

Choobineh, A.; Daneshmandi, H.; Saraj Zadeh Fard, S.Kazem.; Tabatabaee, S.Hamidreza., 2017: Prevalence of Work-related Musculoskeletal Symptoms among Iranian Workforce and Job Groups. Musculoskeletal disorders (MSDs) are known to cause occupational injuries. This study aimed to collate the existed relevant data and develop a general feature of MSDs problem among Iranian workforce. In this study, we used the raw data related to...

Jones, P.; Tate, T., 1988: Emerging technologies for extension delivery systems. Proceedings of the 2nd International Conference on Computers in Agricultural Extension Programs Fedro S Zazueta: 527

Safian, R., D.; Schreiber, T., L.; Baim, D., S., 1993: Specific indications for directional coronary atherectomy: Origin left anterior descending coronary artery and bifurcation lesions. Directional coronary atherectomy is emerging as the treatment of choice for many patients with significant lesions involving left anterior descending coronary artery origin or vessel bifurcations. It offers the potential advantages of safe, reliab...

Bruguera, M.; Gómez-Durán, E.L.; Arrizabalaga, P.; Arimany-Manso, J., 2015: Claims of alleged malpractice. Are there differences between male and female physicians?. Medicina Clinica 143(9): 420-421

Bingle, C.D.; Gorr, S-U., 2004: Host defense in oral and airway epithelia: chromosome 20 contributes a new protein family. The innate immune response is of pivotal importance in defending the mucosal barriers of the body against pathogenic attack. The list of proteins that contribute to this defense mechanism is constantly being updated. In this review we introduce a...

Armon, Y.; Cooper, D.M.; Zanconato, S., 1991: Maturation of ventilatory responses to 1-minute exercise. To test the hypothesis that ventilatory responses to exercise mature during growth in healthy children, we examined CO2 production (VCO2) and minute ventilation (VE) before, during, and for 10 min after 1-min bursts of cycle ergometry exercise. Te...

Li, J.; Gordon, S.C.; Rupp, L.B.; Zhang, T.; Trudeau, S.; Holmberg, S.D.; Moorman, A.C.; Spradling, P.R.; Teshale, E.H.; Boscarino, J.A.; Daida, Y.G.; Schmidt, M.A.; Lu, M., 2016: Long-term progression of viral load and serum markers of fibrosis among treated and untreated patients with chronic hepatitis B. Antiviral therapy for patients with hepatitis B (HBV) infection is generally deferred for "immune inactive" patients, although longitudinal changes in viral load and liver fibrosis remain understudied in this population. Likewise, in tre...

Long, B.J.; Grigoryev, D.N.; Nnane, I.P.; Liu, Y.; Ling, Y.Z.; Brodie, A.M., 1999: In vitro and in vivo inhibition of LNCaP prostate cancer cell growth by novel inhibitors of androgen synthesis. Proceedings of the American Association for Cancer Research Annual Meeting 40: 64, March