Shannon Odelberg Lab
notes modified by REM underlined
- Add 0.05g paraformaldehyde to 75 ml dH2O in a 100 glass beaker, with a small spinbar and heat on stir plate in Hood # 1. (next to the Freezer).
- When it reaches 65°C or begins to bubble add 2-3 drops 1M NaOH and wait until the solution clears. Usually less than 30s.
- Add 20 ml 5X buffer and adjust final volume to 100 ml with dH2O. Aliquot and freeze at -20°C.
5X Buffer 200 ml
- 10 mM sodium periodate = 2.14g
- 100 mM lysine-HCl= 18.27g
- 150 mM NaCl= 1.74g (adjusted for mammals)
- 3 mM KCl=12 ml of 0.25M stock
- 10 μM CaCl2 100 μl of 100mM stock
- 5 mM HEPES = 1.19g
- Check and adjust pH to 7.4