Silver Stained Gels
1. It is strongly recommended that you use coomassie stain rather than silver.
2. If you can not visualize bands with coomassie try to scale up your isolation to increase the amount of protein until you get to coomassie detectable limits.
3. If you can not reach coommassie stainable levels you must use a mass spec compatible silver stain protocol. Only use methanol and acetic acid during the fixing step. Do not use any solutions containing formaldehyde or glutaraldehyde to fix the gel. Refer to Shevchenko et al. (1996) Analytical Chemistry, 68:850-858 for more details.
4. It is better to have all the protein in one band/spot, but if you can not load more protein in one lane then run several lanes and combine the bands/spots.
5. Use 1.0 to 1.5mm gels.
6. Only stain the gel long enough (usually only a few minutes) to detect the bands of interest.
7. Take a picture of the gel and submit it (photocopy or electronic) along with samples.
8. Excise gel band(s)/spot(s) with as little excess empty gel as possible.
9. Place the gel band(s)/spot(s) into a micro centrifuge tube with some ddwater.
10. Fill out Sample Submission Form form.
11. Drop off or send samples and submission form to the facility (Note: No need to send samples on dry ice).