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Services

Services Offered

Protein Identification 1 - Discovery

High sensitivity (silver stain OK), up to 100 proteins or more may be identifiable.
Cost: $200 each
Time: Four to 10 working days

Protein Identification 2 - Confirmation

Coomassie only, up to four proteins may be identifiable.
Intended 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 -- For SRM/MRM methods on our tandem quadrupole MS please inquire!
NEW!! Large-scale PhosphoProteomics! - Please Inquire!!

Breaking News!  Coming soon....  Metabolomics, stay tuned!

The Orbitrap has been installed and is performing well, most qualitative analyses are now run on it!  

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  !!!


© 2014 The University of Texas MD Anderson Cancer Center