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13C). The yields for the first two steps ranged from 70 - 90% while the yields for the last step ranged from 20 - 90%. The purity of each final product was greater than 95% as determined by HPLC/MS analysis
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13C). The yields for the first two steps ranged from 70 - 90% while the yields for the last step ranged from 20 - 90%. The purity of each final product was greater than 95% as determined by HPLC/MS analysis.
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Fluorescence polarization assays were run in 20 μL volume in 384 well format with black plates. The assay solution was 25 mM Hepes at pH 7.5/1 mM tris(2-carboxyethyl)phosphine/0.005% Tween 20 and 20 nM AVPIAQK-rhodamine. XIAP BIR2 was present at 1 μM or XIAP BIR3 at 0.2 μM. Compound was present within a range of 100 μM to 6.1 nM using two fold dilutions starting at 100 μM generating a 16 point curve. Plates were read on an Analyst HT in fluorescence polarization mode with excitation at 530 nm, emission at 580 nm and a dichroic mirror at 565 nm. Resulting data in mP was fit in GraphPad Prism 5 with a non-linear regression using a sigmodial curve with variable slope
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Fluorescence polarization assays were run in 20 μL volume in 384 well format with black plates. The assay solution was 25 mM Hepes at pH 7.5/1 mM tris(2-carboxyethyl)phosphine/0.005% Tween 20 and 20 nM AVPIAQK-rhodamine. XIAP BIR2 was present at 1 μM or XIAP BIR3 at 0.2 μM. Compound was present within a range of 100 μM to 6.1 nM using two fold dilutions starting at 100 μM generating a 16 point curve. Plates were read on an Analyst HT in fluorescence polarization mode with excitation at 530 nm, emission at 580 nm and a dichroic mirror at 565 nm. Resulting data in mP was fit in GraphPad Prism 5 with a non-linear regression using a sigmodial curve with variable slope.
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7444262377
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Z. Nikolovska-Coleska, R. Wang, X. Fang, H. Pan, Y. Tomita, P. Li, P.P. Roller, K. Krajewski, N. Saito, J. Stuckey, and S. Wang Anal. Biochem. 332 2004 261
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6) δ 169.6, 169.3, 132.6, 131.1, 127.4, 125.9, 114.5, 111.1, 110.4, 100.9, 60.2, 56.4, 55.9, 47.3, 38.2, 31.3, 30.0, 29.5, 24.7, 19.0, 18.2, 16.3. MS m/z = 414 (M+H)
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6) δ 169.6, 169.3, 132.6, 131.1, 127.4, 125.9, 114.5, 111.1, 110.4, 100.9, 60.2, 56.4, 55.9, 47.3, 38.2, 31.3, 30.0, 29.5, 24.7, 19.0, 18.2, 16.3. MS m/z = 414 (M+H).
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76449098262
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