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 on selective plates, allow to grow for 1 day. (24°C)
- Place nitrocellulose filters on gel and incubate overnight at 23°C. (Make sure to roll the sheet with a tube)
- Wash filter in water.
- Block filter with BSA for 1 hour.
- Wash in primary antibody overnight at 4C or >3h at room temp
(10,000x dilution)
- Rinse in Tbst.
- Secondary antibody at 10 000X for 45min-1h.
- Rinse in TBST.
- Develop
