Rinse thoroughly with water. Use enough blocking solution to completely cover the filter. B. Place membrane in antibody solution, ensuring that the membrane surface which was in contact with the gel is facing upward. If rinsing more than one filter, make sure to keep filters separate. B. Transfer all filters to a container containing fresh TBST and wash for 5 min.
Repeat wash twice.
Secondary Antibody A. Transfer filters to a clean container. Add secondary antibody diluted in Blocking Solution. Substrate Incubation A. Transfer all filters to container containing substrate solution. B. Follow manufacturer's directions for detection. All trademarks are property of Becton, Dickinson and Company unless otherwise noted.