Bio-Rad Laboratories, Inc.
SEQUENTIAL MULTIPLEX WESTERN BLOTTING

Abstract:

Described are methods and compositions for sequential multiplex detection of target analytes in a sample. The method comprises contacting the sample comprising analytes immobilized on a solid support with binding agents that specifically bind an analyte in the sample, wherein each of the binding agents binds a different analyte and is attached to a single-stranded nucleic acid molecule comprising a unique sequence. The sample is then contacted with a labeled complementary nucleic acid molecule that binds the single-stranded nucleic acid molecule attached to one binding agent. The signal from the label is detected, and then reduced or eliminated. The sample can be simultaneously contacted with a second labeled complementary nucleic acid molecule that binds a different binding agent, and the signal from the second label is detected. The process is repeated for each additional analyte in the sample, thereby sequentially detecting the presence of the analytes in the sample.