Cell Lysates For Western Blotting

Reagents / Solutions

Lysis Buffer:

10ml 10% Sodium dodecyl sulphate (SDS)

10ml Glycerol

10ml b-mercaptoethanol

8ml 0.5M Tris pH6.8

1ml 0.1% bromophenol blue

51ml H2O

Protocol

Spin down 106 cells and wash, if required, in phosphate buffered saline.

Resuspend cells in 100µl warmed lysis buffer, vortex then boil for 5 - 10 minutes to break up DNA.

Cool (not on ice - the SDS will precipitate) and centrifuge to collect droplets.

Vortex briefly to mix.

The samples may now be analysed by , or

Store samples at -20℃ until required.