Preparation of Eukaryotic Lysates for 2D Gel Electrophoresis
This protocol was adapted from “Preparative 2D Gel Electrophoresis with Immobilized pH Gradients,” Chapter 4 in Proteins and Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.INTRODUCTION
Cultured mammalian cell lines are widely used as study models in biomedical research. The protocol described here has been used successfully for many human colon carcinoma cell lines and fibroblast cell lines. Although most cellular proteins can be extracted by this protocol, some DNA-binding proteins may not be recovered. If DNA-binding proteins are to be studied, a nuclease (e.g., Benzonase) should be included. Expression proteomic studies frequently require the detection of low-abundance proteins.
