Making Western Samples Directly from Dicty Cultures

To make gel samples from 24-well plates. Do all harvesting work in hood w/ sterile tools.

  1. Start a boiling water bath
  2. Resuspend cells by pipetting up and down 5-10 times (@400ul)
  3. Spin down the cells in microfuge set to 3k rpm for 2min.
  4. Aspirate off supe, respin for 1 min @ 3k rpm. Remove remaining supe
  5. Add 40ul of TE, quickly mix and put in boiling water bath immediately for 5 min.
  6. Add 40ul of 2x SDS SB and boil for 5min.
  7. Load 10ul on gel