Difference between revisions of "CPY Assay"
From Bridges Lab Protocols
(Created page with '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 sele...') |
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*Primary antibody (polyclonal against cpy location?) how long? | *Primary antibody (polyclonal against cpy location?) how long? | ||
*Secondary antibody (where/what is it and how long?) | *Secondary antibody (where/what is it and how long?) | ||
| + | Wash with pbs in between? | ||
| + | |||
| + | What is ECL? | ||
Revision as of 16:38, 7 July 2010
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?
