Cell Lysis

1. Aspirate media.
2. Add 500 uL of cold PBS to each well.
3. Aspirate.
4. Add 200 uL of trypsin.
5. Incubate for 90 seconds.
6. Neutralize with 800 uL of cold complete media.
7. Pipette up and down to disperse clumps.
8. Transfer contents of each well to 2 mL labelled eppendorf tubes.
9. Centrifuge/aspirate.  How long?  How fast?
10. Add lysis buffer.  How much?
11. Incubate at 4C for 30 minutes with constant agitation.
12. Centrifuge at 12000 rpm for 20 minutes.  Long/fast?
13. Remove tubes; place on ice.
14. Transfer supernatant to a new tube (on ice).  *At this point you can centrifuge to concentrate if necessary. - protocol?

Bradford Assay (to check protein concentration)

1. Fill this in!