Protein Mass Spectrometry Identification

The identification/analysis of protein spectra of single proteins or protein mixtures, as well as the protein sequence analysis of single proteins, are common issues encountered in biological research. Sequence information obtained through mass spectrometry for protein sequencing and protein mass spectrometry identification enables protein identification, i.e., protein identification based on LC-MS. Common analyses include:Mass spectrometry determination of protein molecular weight,N-terminal and C-terminal sequencing of proteins,Protein sequence analysis and,Post-translational modification site analysis, etc..

BiotechPack uses Thermo's latest Obitrap Fusion Lumos mass spectrometer combined with Nano-LC nanoliter chromatography technology to provide efficient and accurate protein mass spectrometry identification services for protein extracts, SDS-PAGE protein bands, 2D protein spots, pull-down, and Co-IP samples.

BiotechPack offers the following protein mass spectrometry identification services:

1. Protein mass spectrometry analysis/identification for purified proteins;
2. Mass spectrometry identification of protein gel spots, protein gel strips, silver-stained gel mass spectrometry identification, and identification of gel spots/strips stained with Coomassie/fluorescence/Zn/Cu2+, etc.; refer to:Protein gel spot, gel strip, IP sample protein identification services;
3. Protein identification/analysis for pull-down protein mass spectrometry identification, IP protein interaction IP-MS interaction protein identification, Co-IP mass spectrometry validation interaction proteins, immunoprecipitation combined with mass spectrometry validation in simple mixture samples;
4. Protein mass spectrometry analysis/identification of unknown proteins in complex samples. Even very low concentration protein samples may be identified through protein mass spectrometry.

For the identification of single proteins and simple protein mixtures, we guarantee a 100% identification rate, otherwise no fees will be charged.

NanoLC-MS/MS analysis of proteins in simple or complex mixtures. Samples can be from SDS-PAGE gel bands or protein spots, or solution samples.(We also provide 1D or 2D SDS-PAGE services, click to learn more.)

BiotechPack mass spectrometry-based protein identification examples

BiotechPack mass spectrometry-based protein mass spectrometry identification process:

Sample pretreatment
First, reduce the protein sample with dithiothreitol (DTT) and alkylate with iodoacetamide (IAA). Then digest the sample with trypsin, generally resulting in cleavage after lysine and arginine residues. Finally, lyophilize to obtain peptide fragments.

Instrument analysis
Dissolve the sample in NanoLC's phase A before loading, and analyze under the NanoLC-MS/MS system.

Database search
Download the corresponding species proteome database from UniProt based on the sample's species origin. Use software like MaxQuant, Proteome Discoverer, or Peaks to match the data obtained from liquid chromatography-mass spectrometry with the database data, identifying proteins in the sample through the mass of primary and secondary peptide fragments.

If the sample's species does not have a complete proteome database, we will use the proteome database of a closely related species or the same model organism for protein identification analysis.

Report issuance
The identification report will include detailed sample pretreatment steps, chromatography-mass spectrometry platform parameters, and a list of identified proteins and peptides. Raw mass spectrometry data will be provided via a cloud storage link.

You only need to collect and provide your samples to us, and all subsequent steps will be completed by us.BiotechPack's mass spectrometry-based protein mass spectrometry identification/analysis services include: Sample pretreatment - Protein reduction and alkylation followed by digestion with highly specific proteases to obtain peptide fragments; Instrument analysis - Separation of peptides at the highest sensitivity using nano-flow chromatography combined with high-resolution mass spectrometry, collecting peptide mass spectrometry data using ultra-high-resolution mass spectrometers; Database search - Analysis of thousands of mass spectra using the sample's species-specific protein database (using the Uniprot database, updated monthly to ensure access to the latest data), achieving protein mass spectrometry identification in the sample; culminating in the issuance of a sample report, all completed by BiotechPack in a one-stop manner:

1. The client provides samples containing one or more proteins, such as gel strips, gel bands, solution or solid samples, pull-down or Co-IP samples.
2. BiotechPack selects the appropriate enzyme based on the sample's condition for protease digestion, followed by nanoLC-MS/MS analysis.
3. Conduct database search analysis on the obtained data.
4. Receive the report within 10-15 working days.

*If your sample contains detergents/lysis agents, please indicate on the sample submission form.