Recipes O

2x PK buffer

100 mM KPO4 20 mM MgCl2 10 mM NaF

9 mM dithiothreitol

GST Interaction buffer

10 mM MgCl2

1 mM dithiothreitol 0.1% Nonidet P-40 1% nonfat dry milk

GST Wash buffer 1

Phosphate-buffered saline containing 0.2% Triton X-100. GST Wash buffer 2

Phosphate-buffered saline containing 0.2% Triton X-100 and 100 mM KCl.

GST basic buffer

10 mM MgCl2 1 mM dithiothreitol 0.1% Nonidet P-40

GST blocking buffer

1 mM dithiothreitol 0.1% Nonidet P-40 5% nonfat dry milk

Dissolve an appropriate amount of solid KCl in H2O, autoclave for 20 minutes on liquid cycle and store at room temperature. Ideally, this 4 M solution should be divided into small (approx. 100 pl) aliquots in sterile tubes and each aliquot thereafter used one time.

NaCl

To prepare a 5 M solution: Dissolve 292 g of NaCl in 800 ml of H2O. Adjust the volume to 1 liter with H2O. Dispense into aliquots and sterilize by autoclaving. Store the NaCl solution at room temperature.

137 mM NaCl 2.7 mM KCl 10 mM Na2HPO4

2 mM KH2PO4

(Phosphate-buffered Saline) Dissolve 8 g of NaCl, 0.2 g of KCl, 1.44 g of Na2HPO4, and 0.24 g of KH2PO4 in 800 ml of distilled H2O. Adjust the pH to 7.4 with HCl. Add H2O to 1 liter. Dispense the solution into aliquots and sterilize them by autoclaving for 20 minutes at 15 psi (1.05 kg/cm 2) on liquid cycle or by filter sterilization. Store the buffer at room temperature. If necessary, PBS may be supplemented with 1 mM CaCl2 and 0.5 mM MgCl2.

Was this article helpful?

0 0

Post a comment