B. Hydrolysis 1. Incubate at 70C for 10 minutes 2. Neutralize by addition of 15 m L 0.1 N HCl. Add 450 m L water to each reaction. Spin at 12g for 7 minutes.
Discard flow through.
Repeat process two more times, refilling original filter with 450 m L water. 10 m L. Samples can be stored at -20C indefinitely. 9.0 to 50 mM final. Each tube is sufficient to label 10 reactions under normal conditions. Aliquot 2 m L into 10 single use tubes that are then dried in vacuo and store desiccated at 4C. NHS-ester conjugated Cy dye is rapidly hydrolyzed in water, therefore, do not store in DMSO or water.