In-Gel Proteolytic Digestion & Mass Spectrometric Analysis

 

Protein samples can be subjected to either one- or two-dimensional SDS-polyacrylamide gel electrophoresis (SDS-PAGE). After staining, the bands (1D SDS-PAGE) or spots (2D SDS-PAGE) are excised, alkylated and then digested with trypsin. The resulting peptides are then analyzed by LC/MS/MS, nanospray ESI--MS/MS or MALDI-Tof-MS. MALDI-MS and nanospray are the most sensitive. These systems are capable of analysis of low femtomole quantities of peptides. The most complete analysis is obtained by LC/MS/MS analysis utilizing a 75 micron ID x 150mm reversed-phase-HPLC column. The sensitivity of this method is currently limited to ~50-100 femtomoles of protein digest. The excised gel band(s) containing the protein of interest should be submitted in a microcentrifuge tube containing ~10µl of 20% methanol/water. Alternately, the complete stained gel can be submitted along with a map indicating the spots to be treated/ analyzed, and we will excise the spots.

Prices

Sample Prep
In-gel alkylation & tryptic digestion (single sample):
$150
Additional samples in set:
$60
Analysis Options
Best: LC/MS/MS with Mascot database search
$115
Good: Nanospray-MS/MS  with Mascot database search
 $75
Economy: MALDI-MS  with Mascot database search 
$60

Multiple sample sets qualify for additional discounts for both sample prep and analysis charges