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Volumn 276, Issue 5315, 1997, Pages 1125-1128

Engineering chemical reactivity on cell surfaces through oligosaccharide biosynthesis

Author keywords

[No Author keywords available]

Indexed keywords

AVIDIN; HYDRAZIDE; KETONE; MANNOSAMINE; OLIGOSACCHARIDE; RICIN A;

EID: 0030929377     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.276.5315.1125     Document Type: Article
Times cited : (660)

References (31)
  • 15
    • 1842363146 scopus 로고    scopus 로고
    • note
    • 3, 0.1% NCS in PBS, pH 7.4). The cells were then suspended in 100 μl of FITC-avidin-staining solution (of FITC-avidin at 5.6 μg/ml in avidin-staining buffer). After a 10-min incubation in the dark at 0°C, the cells were diluted with 2 ml of cold avidin staining buffer and washed twice. The cells were resuspended in 400 μl of avidin staining buffer and subjected to flow cytometry analysis.
  • 17
    • 1842263775 scopus 로고    scopus 로고
    • note
    • 7 cells/ml. Cells were prepared for flow cytometry analysis as described in (15).
  • 22
    • 1842269599 scopus 로고    scopus 로고
    • note
    • 120 for 15 min, washed twice with PBS (pH 7.4), and analyzed by flow cytometry.
  • 23
    • 1842394942 scopus 로고    scopus 로고
    • note
    • The relation between fluorescence intensity observed by flow cytometry analysis and the number of fluorescent molecules per cell were determined with biotinylated polystyrene beads (Spherotech) with a predetermined number of biotin molecules per bead and with a similar diameter to that of Jurkat cells.
  • 24
    • 0025093858 scopus 로고
    • and references therein
    • S. Sell, Human Pathol. 21, 1003 (1990), and references therein.
    • (1990) Human Pathol. , vol.21 , pp. 1003
    • Sell, S.1
  • 28
    • 1842312921 scopus 로고    scopus 로고
    • note
    • 2 groups of lysine residues on avidin were modified to present pyridyldithio groups by reaction with N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) (Pierce) as follows. A solution of egg-white avidin (2 mg, Sigma) and SPDP (4 μl of a 250-mM solution in dimethyl sulfoxide) was incubated at room temperature for 1 hour. Excess SPDP was removed by Sephadex G-25 gel filtration and the modified avidin was collected into PBS buffer (pH 7.4). RTA-2-IT was added to this solution in a 1:1 molar ratio and incubated at room temperature for 18 hours. The 1:1 RTA-avidin conjugate was purified from the reaction by Sephadex G-150 gel filtration and characterized by non-reducing and reducing SDS-polyacrylamide gel electrophoresis.
  • 31
    • 1842321508 scopus 로고    scopus 로고
    • note
    • We thank A. Fischer, P. Schow, and P. Giblin for stimulating discussions and technical advice. Supported by an ARCS Foundation graduate fellowship (L.K.M.) and by grants from the Camille and Henry Dreyfus Foundation, the Pew Scholars Program, the Alfred P. Sloan Foundation, the Exxon Education Foundation, the W. M. Keck Foundation, and the Laboratory-Directed Research and Development Program of Lawrence Berkeley National Laboratory under the Department of Energy, contract number DE-AC03-76SF00098 (C.R.B.).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.