Profiling pancreatic cancer-secreted proteome using 15N amino acids and serum-free media.

ABSTRACT:

Objectives

A new method of determining protein turnover by labeling protein with N amino acids was used in conjunction with serum-free cell culture to profile secreted proteins that are released by MIA PaCa-2 pancreatic cancer cells in culture.

Methods

MIA PaCa-2 cells were first cultured in Dulbecco modified Eagle medium (Gibco by Invitrogen, Carlsbad, Calif) with 10% fetal bovine serum, then in serum-free modified Eagle medium with or without 50% N algal amino acid mixture. The effect of oxythiamine chloride on secreteome was studied. Secreteome from cell culture media was analyzed by 2-dimensional (2D) gel electrophoresis. Differentially expressed proteins were detected and identified. Protein turnover rates were calculated according to the newly established method. Weste

SUBMITTER: Xiao J 

PROVIDER: S-EPMC2835986 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature