CPY Assay
From Bridges Lab Protocols
Carboxypeptidase Secretion Assay
Protocol
- Revive Yeast Cells (2-3) days
- Pick one colony and make a 1/2 inch x 1/2 inch square patch of yeast. (Does this have to be selective plates? or are YPD plates always used?)
- Place nitrocellulose filters on gel and incubate overnight at 23(So you're not actually moving the patch to the filter, but just allowing it to touch?)
- Wash filter in water (by adding water to dish and rocking?)
- Primary antibody (polyclonal against cpy location?) how long?
- Secondary antibody (where/what is it and how long?)
Wash with pbs in between?
What is ECL?