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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!!

Metabolomics  -  Please email (or call) for an appointment to discuss your project!

The brand new Orbitrap Fusion has been installed and is performing well!  We gratefully acknowledge the NIH High-End Instrumentation program grant# 1S10OD012304-01 for funding our acquisition of the newest Orbitrap technology available today.

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