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2
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0029140496
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L. E. Canne, A. R. Ferré-D'Amaré, S. K. Burley, S. B. H. Kent, ibid. 117, 2998 (1995).
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Canne, L.E.1
Ferré-D'Amaré, A.R.2
Burley, S.K.3
Kent, S.B.H.4
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3
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0025117445
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D. Rideout, T. Calogeropoulou, J. Jaworski, M. McCarthy, Biopolymers 29, 247 (1990).
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(1990)
Biopolymers
, vol.29
, pp. 247
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Rideout, D.1
Calogeropoulou, T.2
Jaworski, J.3
McCarthy, M.4
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10
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0000792452
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C.-H. Lin, T. Sugai, R. L. Halcomb, Y. Ichikawa, C.-H. Wong, J. Am. Chem. Soc. 114, 10138 (1992).
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(1992)
J. Am. Chem. Soc.
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, pp. 10138
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Lin, C.-H.1
Sugai, T.2
Halcomb, R.L.3
Ichikawa, Y.4
Wong, C.-H.5
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15
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1842363146
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note
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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.
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17
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1842263775
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note
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7 cells/ml. Cells were prepared for flow cytometry analysis as described in (15).
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22
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1842269599
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note
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120 for 15 min, washed twice with PBS (pH 7.4), and analyzed by flow cytometry.
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-
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23
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1842394942
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note
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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.
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24
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0025093858
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and references therein
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S. Sell, Human Pathol. 21, 1003 (1990), and references therein.
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(1990)
Human Pathol.
, vol.21
, pp. 1003
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Sell, S.1
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27
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0021891359
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A. J. Cumber, J. A. Forrester, B. M. J. Foxwell, W. C. J. Ross, P. E. Thorpe, Methods Enzymol. 112, 207 (1985).
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(1985)
Methods Enzymol.
, vol.112
, pp. 207
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Cumber, A.J.1
Forrester, J.A.2
Foxwell, B.M.J.3
Ross, W.C.J.4
Thorpe, P.E.5
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28
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1842312921
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-
note
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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.
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31
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1842321508
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note
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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.).
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