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Optimal Conditions in 384-well plate: Glucose oxidase (4 U/mL), maltose phosphorylase (2 U/mL), maltose (0.2 mM), amplex red (50 μM), horseradish peroxidase (0.2 U/mL), ATP (1.5 mM), and Hsp90 (50 μg/mL) were combined to total of 50 μL and incubated for 3 h at 37 °C and the absorbance of the resorufin product was measured at 563 nm.
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Control conditions: Glucose oxidase (4 U/mL), maltose phosphorylase (2 U/mL), maltose (0.2 mM), amplex red (50 μM), horseradish peroxidase (0.2 U/mL) Hsp90 (50 μg/mL), compound (20 μg/mL), and inorganic phosphate (100 μM) were combined and incubated for 3 h at 37 °C and the absorbance of resorufin was measured at 563 nm.
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