> Western Blotting
> Western Blot Buffers
Transfer Buffers 5X transfer buffer(-MeOH)
500mL
1L
Tris base
9.37g
18.74g
Glycine
45.0g
90.0g
Blocking Buffer
10X (skim milk)
1 litre
140mM NaCl
1.4M NaCl
81.8g
10mM KPO4 (pH 7.4)
0.1M KPO4
200mL of 0.5M stock
0.02% Azide
0.2% azide
2g/litre
(0.5 % skim milk powder 5g/litre)
AB Buffer
Polyclonal Ab
1 litre wash buffer plus BSA
1 litre wash buffer plus ( )
0.1% Triton
560mM NaCl (+ BSA 10g)
10mM KPO4
0.02% SDS (+ 1mL 20% SDS)
0.02% azide
10mM KPO4 (+ 24g NaCl)
1% BSA
0.01% azide
Wash Buffer
10X (1 litre)
80g
0.02% azide NaN3
2g
10mL
WESTERN AP BUFFER 500 ml 1L 2L
2L
100mM Tris pH 9.5
6g
12g
24g
100m NaCl
12.5mL 4M stock
25mL
50mL
stock (2.92g)
(11.68g of solid)
5mM MgCl2
0.51g
1.02g
2.04g
stock(2.92 g) (11.68g of solid)
* Add about 350 ml H2O to dissolve - Adjust pH to 9.5 with HCl make to 500 ml final volume with water. *