Proteomics is a contemporary and throughput technology analyzing the entire complement of proteins in a biological system. The featured characterization is a holistic examination of all the interweaving biological pathways and networks. It has been an indispensable tool in molecular and cellular biology, biomedicine, and biodefence as well as in the emerging field of systems biology. The technique relies on specialized liquid chromatography-tandem mass spectrometry (LC-MS/MS) to achieve accurate, highly sensitive, and rapid sequencing and quantification of thousands and millions of peptides to infer proteins and their structural modifications. The acquired data relies on biostatistics and bioinformatics for data mining and pattern recognition to gain biological insights.

In this workshop, participants will be able to learn the fundamental theory of this widely used technology in one hour tutorial, and subsequently work in the lab for the rest of the day to practice a well-designed complete proteomic procedure outlined in the workshop program below including sample preparation, instrumentation, and data analysis. Two parallel modules are offered at this time with one focusing on general proteomics and the other on membrane proteomics. It is recommended that for beginners, take the general proteomics module first as the membrane proteomics module is designed for trainees already equipped with basic proteomics knowledge.

Module 1 (General Proteomics)

Starting material of the workshop: Proteins of 100ug in powder form or in buffer solution with known composition will be needed for this workshop. Attendees are encouraged to bring their own samples, otherwise we will provide sample proteins for participants to practice.

Itinerary of the day: 9:00am Presentation of the basic principle of the methodology, the key steps, and the general application of the methodology.10:00am Bench work: Denaturation, alkylation, quenching, enzymatic digestion.12:00am Group lunch.1:00pm SPE cleaning up and speedvac concentration.3:00pm Discussion of this method.4:00pm Nanodrop, MS characterization and database search and analysis5:00pm Wrap up.

Module 2 (Membrane Proteomics)

Starting material of the workshop: Clean and dried 100ug tryptic membrane peptides are needed for this workshop. Attendees are encouraged to bring their own sample peptides, otherwise we will provide sample peptides for trainees to practice.

Itinerary of the day: 9:00am Presentation of the basic principle of the methodology, the key steps, and the general application of the methodology.10:00am Bench work: Dispersion, oxidation, coupling of peptides to the resin.12:00am Group lunch.1:00pm Wash and release the captured peptides.3:00pm Discussion of this method.4:00pm Nanodrop, MS characterization and database search and analysis5:00pm Wrap up.