Dot blot - glasgowlab/home GitHub Wiki

  1. boil 95C 5min (do not use loading dye)
  2. spot 1-2ul onto nitrocellulose membrane
  3. dry for 15min
  4. block 1 hr 5% milk in tbst
  5. add primary ab (anti His mouse, fridge drawer) to the milk at 1:5000 ratio, rotate 1 hr
  6. wash 3x for 5 min in TBST
  7. add secondary ab (peroxidase anti mouse IgG) in TBST at 1:5000 ratio, rotate 1 hr
  8. wash 3x for 5 min in TBST
  9. 1ml each of both chemiluminescent substrates
  10. image