Protein or Peptide Gel based identification

Identification by MALDI-MS/MS

MALDI mass spectrometry is a fast and robust technique where the mixture of peptides is deposited in a dry matrix and analyzed by MS and MS/MS. This analysis is best suited for bands or spots from 1D/2D gels containing 1-3 proteins where the peptide mixture is not too complex.   

Identification by nanoLC-MS/MS

Nano-LC-MS/MS analysis is a slow, delicate and very sensitive technique where the mixture of peptides is separated by a HPLC gradient and each peptide analyzed in-solution by ESI MS/MS on a QTOF mass spectrometer. This analysis is well-suited for complex mixtures of peptides from multiple proteins, and often identifies 5-50 proteins in each sample.  

Identification service by both MALDI MS/MS and nanoLC MS/MS gives you:

 The highest sensitivity - Coomassie & Silver stained proteins are identified.  Fast analysis - Identification results.  High confidence - Identification based on MS/MS peptide sequencing.  Documentation - Clear and easy-to-understand reports.
<div style="\&quot;text-align:" center;\"="">Sample Preparation Guidelines Proteins from 1D SDS PAGE and 2D PAGE gels can be submitted for protein identification analysis.

1. Run your 1D/2D gel. 2. Stain the gel with Coomassie Blue, SYPRO Ruby or Silver stain

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1. Coomassie Blue and SYPRO Ruby works well in combination with mass spectrometry

2. Silver stain –Do not destain silver-stained samples!

3. Cut out the bands/spots of interest 1. Use a clean scalpel and tweezers 2. Cut out the gel band close to the stain edges to have the highest protein concentration in the smallest gel volume. Ideal band size is 2 x 5 mm.  Do not cut the gel sample into tiny, tiny gel pieces! 3. Clean the scalpel in-between spots/bands

4. Put the samples into separate wells/tubes with water 1. Use the 96-well plate or Eppendorf Safe  lock tube or similar tubes from your lab. 2. Add 100-200 μl distilled water to keep each gel plug wet. 3. Use the rubber lid for the 96-well plate. Make sure the lid is properly closed.

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