Mass Spectrometry-Based Protein Identification Service

The identification and analysis of single proteins or protein mixtures via mass spectrometry, as well as the analysis of individual protein sequences, are critical tasks in biological research. Techniques include determining protein molecular weights, sequencing protein N- and C-termini, analyzing protein sequences, and identifying post-translational modification sites.

 

MtoZ Biolabs utilizes Thermo's latest Orbitrap Fusion Lumos mass spectrometer in conjunction with Nano-LC technology. This equipment allows for the precise and efficient identification of proteins from various samples such as protein extracts, SDS-PAGE bands, 2D gel spots, pull-down, and Co-IP assays.

 

Services at MtoZ Biolabs

1. Mass Spectrometry Analysis for Purified Protein

2. Mass Spectrometry of Proteins from Gel Spots and Strips, Including those with Silver, Coomassie Brilliant Blue, Fluorescence, and Zn/Cu2+ Staining

3. Analysis and Identification of Proteins in Simple Mixtures after Pull-Down, IP, and Co-IP by Mass Spectrometry

4. Analysis and Identification of Unknown Proteins in Complex Samples, Ensuring Detection even at Very Low Concentrations

 

We guarantee a 100% identification rate for single proteins and simple protein mixtures, with no charge if identification is unsuccessful.

 

Analysis Workflow

1. Sample Preprocessing

Protein samples are first reduced with dithiothreitol (DTT) and alkylated with iodoacetamide (IAA). Trypsin digestion follows, typically cleaving at lysine and arginine residues. The samples are then lyophilized to obtain peptide fragments.

 

2. Machine Analysis

The samples are redissolved in the mobile phase A of NanoLC and analyzed using the NanoLC-MS/MS system.

 

3. Database Search

For each sample, the corresponding species' proteome database is downloaded from UniProt. Tools such as MaxQuant, Proteome Discoverer, and Peaks are used to match the mass spectrometry data with database entries, identifying proteins by comparing the masses of primary and secondary peptide fragments.

 

If a complete proteome database for the sample's species is not available, databases from closely related species or model organisms are used for analysis.

 

4. Issuing a Report

The final report includes detailed descriptions of sample preprocessing, LC-MS settings, and lists of identified proteins and peptides. Raw mass spectrometry data is made available via cloud storage.

 

You only need to collect your samples and provide them to us, and we will complete all subsequent steps. MtoZ Biolabs' mass spectrometry-based protein mass spectrometry identification/analysis service includes: sample preprocessing - protein reduction alkylation followed by digestion with highly specific proteases to obtain peptide fragments; machine analysis - separation of peptides through nano-flow chromatography combined with high-resolution mass spectrometry, collecting peptide mass spectrometry data at the highest sensitivity; database search - analyzing thousands of mass spectra (using the Uniprot database, updating the database monthly to ensure access to the latest data), thus achieving protein mass spectrometry identification of the sample; finally issuing a sample report, all done by MtoZ Biolabs in a one-stop manner:

 

(1) Provides a sample containing one or more proteins, such as gel strips, gel tapes, solutions, solid samples, pull-down, or Co-IP samples.

(2) Based on the sample situation, selects the appropriate enzyme for protein digestion and then performs nanoLC-MS/MS analysis.

(3) Analyze the obtained data.

(4) Receive the report within 10-15 working days.

 

*If your sample contains detergents/solvents, please note on the sample submission form.