Lysis of Cultured Animal Cells, Yeast, and Bacteria for Immunoblotting
Adapted from Proteins and Proteomics, by Richard J. Simpson. CSHL Press, Cold Spring Harbor, NY, USA, 2003.INTRODUCTION
This protocol describes cell lysis for immunoblotting. Cell lysis with detergents is commonly used with cultured animal cells. Typically, a 2% solution of the ionic detergent sodium dodecyl sulfate (SDS) is sufficient for lysing cells for the purpose of immunoblotting studies. Both cultured animal cells and bacteria such as Escherichia coli can be lysed in this manner. If the antigenic determinant recognized by the antibody being studied is dependent on the native spatial conformation and sensitive to reducing conditions, then dithiothreitol (DTT) should be omitted from the lysis buffer and non-reducing/non-urea gels might need to be used.
