A Transient Expression Assay Using Arabidopsis Mesophyll Protoplasts

1. Arabidopsis (Ecotype: Columbia) 2. Fetal bovine serum (FBS) (Sigma-Aldrich, catalog number: F6178) 3. PEG4000 (Fluka, catalog number: 81240) 4. Mannitol 5. NaCl 6. KCl 7. CaCl2 8. MgCl2 9. 0.5 M MES (pH 5.7) 10. β-mercaptoethanol 11. Cellulase R10 purchased from http://www.yakult.co.jp/ypi/english/frame03.html 12. Macerozyme R10 purchased from http://www.yakult.co.jp/ypi/english/frame03.html 13. Bright-Line/Dark-Line Counting Chambers (catalog numbers: 3100, 3110, 3200, 3500, 1490, 1492, 1475 and 1483) 14. Enzyme solution (see Recipes) 15. PEG solution (see Recipes) 16. W5 solution (see Recipes) 17. MMg solution (see Recipes) 18. Washing and incubation solution (see Recipes)