Preparation of Blots for Western Blot screening

1. Cast 4 Mini-Preparation SDS-PAGE Gel (one small well and one big well)
2. Load MW marker in the small well
3. Load 0.5-1 microgram of purified protein or 20-50 microgram of crude cell lysates (containing the screening protein) in the large "Preparation Well"
4. Perform electrophoresis
5. Transfer proteins to either nitrocellulose or PVDF membranes
6. Wash membranes twice with distilled water and detect proteins by Ponceau Red staining
7. Soak membranes in PBS with 0.05% sodium azide for 1 minute
8. Seal membranes in plastic bags individually and send them to GENEWIZ at either 4oC or room temperature