Services
Services Offered
Protein Identification 1 - Discovery
High sensitivity (silver stain OK), up to 100 proteins may be identifiable.
NEW COST REDUCTION!! Cost: $200 each
Time: Four to 10 working days
Protein Identification 2 - Confirmation
Coomassie only, up to four proteins may be identifiable.
Recommended for confirmation of purified proteins, such as recombinants
Cost: $100 ea
Time: Three to six working days
Molecular Weight Determination
Check the MW of your peptide or molecule. Proteins may also be measured if enough clean protein is available - but larger proteins are less reliable.
Cost: $50/sample
Time: Four to 10 working days
Quantitative Protein Analysis
Using stable isotope methods, iTRAQ, etc. -- Please inquire.
Using non-isotopic methods -- Our new tandem quadrupole MS has been installed!
SRM/MRM quantitative methodology - Please Inquire!!
Breaking News! The Orbitrap-XL has been installed! Please check with us if your project requires this system!
Do you need higher resolution than you see here or some other MS service?
Please inquire - we are happy to consult with you and help arrange the MS resource you require. PTM analysis? - please inquire.
Required Forms
Proteomics Facility Sample Submission Form (doc) - all users
IDT Authorization for Proteomics Facility (doc) - internal users only
Credit Card Authorization Form (doc) - external users (PO can also be used)
Bio-specimen Transportation Form - for the Employee Shuttle
Shuttle Form (pdf)
Recommended Silver Stain
Download Silver Stain Protocol (pdf). Please ignore destain, we destain after cutting the bands.
Silver Staining of Protein Gels
Reference: Morrissey, J.H. (1981).Silver stain for proteins in polyacrylamide gels: A modified procedure with enhanced uniform sensitivity. Anal. Biochem. 117: 307-310.
Reagents
Silver Solution
- 0.1% AgNO3 in H2O: 0.1g AgNO3 dissolved in 100 ml H2O
Developing Solution
- 3% Sodium carbonate (Na2CO3) in H2O
- 50 µl formaldehyde/100 ml
- Stir for 2 minutes
Procedure
All incubations are at room temperature, and carried out with gentle shaking on a shaking platform. All steps take place in a clean (acid washed) pyrex dish.
Make up all solutions fresh, just before use.
- Incubate 15 min in 50% methanol
- Incubate 15 min in 32 µM DTT (8 µl 1M DTT /250 ml water) solution (freshly made)
- Wash briefly (approximately 5 to 10 seconds) with H2O, twice
- Wash with a little Silver Solution, pour away, then add the rest of the solution and incubate for 15 minutes
- Wash briefly (approximately 5 to 10 seconds) with H2O, twice
- Twice wash with a little Developing Solution, pour away, then add the rest of the solution and incubate until the bands are the desired intensity
- Pour off most of the developing solution, sprinkle solid citric acid into the solution containing the gel and swirl. Continue slowly adding the citric acid until the fizzing ceases
- Add a little H2O, and incubate 15 minutes
- Incubate 3 X 15 minutes with 3 changes of H2O
- Preferred: Bring the gel to our lab; we prefer to cut the bands here
Results from Proteomics Tracker
Now available on the WWW at https://biostatistics.mdanderson.org/ProteomicsTracker !!!