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31344478871
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note
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3: C, 56.54; H, 4.03; N, 19.78. Found: C, 56.31; H, 3.88, N, 19.56.
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14
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31344478083
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note
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2, 2 mM DTT, and 1 mg/ml BSA) containing 0.5 mmol pGAT-biotin and 3-4 ng KDR enzyme was added to each well. After 5-10 min preincubation, the kinase reaction was initiated by the addition of 10 μl of 10 μM ATP in the reaction buffer, after which the plate was incubated at room temperature for 45 min. The reaction was stopped by addition of 50 μl KF buffer (50 mM Hepes, pH 7.5, 0.5 M KF, and 1 mg/ml BSA) containing 100 mM EDTA and 0.36 μg/ml PY20K (Eu-cryptate labeled anti-phosphotyrosine antibody, CIS Bio International) was added and after an additional 2 h incubation at rt, the plate was analyzed in a RUBYstar HTRF Reader.
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15
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0033103867
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M.A. McTigue, J.A. Wickersham, C. Pinko, R.E. Showalter, C.V. Parast, A. Tempczyk-Russell, M.R. Gehring, B. Mroczkowski, C.C. Kan, J.E. Villafranca, and K. Appelt Structure 7 1999 319
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Mroczkowski, B.8
Kan, C.C.9
Villafranca, J.E.10
Appelt, K.11
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16
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0033954256
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H.M. Berman, J. Westbrook, Z. Feng, G. Gilliland, T.N. Bhat, H. Weissig, I.N. Shindyalov, and P.E. Bourne The Protein Data Bank, Nucleic Acids Res. 28 2000 235
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Shindyalov, I.N.7
Bourne, P.E.8
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21
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31344464232
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note
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4, and absorbance was measured at 450 nm.
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