Secretome project - Summer 09
Secretome project - Summer 09
1- Pellet Chlorella (low speed, ~4.5K x 5 min), resuspend half cells in 10 ml BBM, the other half in 10 ml MBBM. Incubate overnight.
2- Pellet cells (4 min at 7K). Save supernatants (Secretome).
3- Wash pellets 1x with 5ml pH 7 MOPS. Centrifuge 4min at 7K. Discard supernatant.
4- Suspend pellets in 1 ml pH 7 MOPS, add 0.11ml 10M LiCl, swirl for 1 min.
5- Centrifuge 4 min at 7K. Withdraw and save supernatants. (Wash)
6- TCA ppt Secretome and Wash samples.
7- Run SDS gels on samples and stain with silver.