Protein running, transfer and blotting buffer

For Western Blotting

Running Buffer 10X.

 

35 mM SDS                (10.08 g)

250 mM Tris               (30.3 g)

1,92 M glycine             (144g)

 

In one liter volume. Dilute to 1X before use.

 

Transfer buffer

 

25 mM Tris                 (3.03 g)

192 mM glycine           (14.4g)

In 800 mL final volume of water. Add 200 mL of methanol 100%. Keep at 4C.

 

Antibody wash buffer (make fresh)

 

TBS-T.

20 mM            Tris pH 7,5      (20 mL of a 1L solution)

150 mM          NaCl                (30 mL of a 5M solution)        

0.02 % Tween 20

 

Antibody incubation/hybridization buffer (make fresh)

 

TBS-T + 2.5% defatted dry milk.