Sample Submission Guidelines
When submitting samples for analysis, keep in mind that sample preparation strongly affects data quality. Take a look at the guidelines below, and if you have any questions, don't hesitate to contact us.
For submission of gel pieces for LC-MS:
Take special care to avoid keratin contamination: always use clean nitrile gloves, dishes, and implements when handling protein gels. Keep the work space and tools dust-free and avoid containers that were previously in contact with western blot blocking reagents.
After cutting out the bands of interest with a fresh razor blade, place the gel pieces in low-binding Eppendorf tubes (e.g. polypropylene tubes).
Notify us whether the gel was stained and if so, what stain was used (Coomassie, fluorescent, silver, etc.) Complete destaining is usually not necessary.
If you plan to silver stain your gel, contact the Protein Sciences Facility first to ensure that your staining protocol is MS-compatible.
For samples submitted for intact mass analysis:
It is best if the samples are submitted as dry material or concentrated liquid in a small low-binding polypropylene Eppendorf tube to maximize recovery.
Ensure that protein samples are as free from salts, detergents, and non-volatile components as possible.
-Brij-35, Triton X-100, CHAPS
-DMF and DMSO
-Tween-20, glycerol, and SDS
-Salts and Tris/phosphate buffers >100 mM
-Formic and acetic acid
-Methanol, ethanol, acetonitrile
For all samples submitted for LC-MS:
Communicate the exact buffer composition. Common biological buffers (phosphate, Tris, HEPES) are acceptable, but many detergents are not MS-compatible and will thus need to be removed before the sample can be analyzed.
Avoid contaminants by using high purity reagents and freshly made solutions and always wearing gloves when preparing samples.
Provide a protein amount and/or concentration.
Tell us what organism(s) the samples are from so we can pick the best databases for analysis. We can also create a custom database as long as the sequence(s) of interest are provided beforehand.
When dropping off samples for analysis, bring a completed sample request form. For users in the University of Illinois system, be sure to include an account number. For off-campus users, complete the same form but contact the facility to provide payment information.