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RECIPES > WESTERN BLOTTING

> Western Blotting

> Western Blot Buffers

 

WESTERN BLOT BUFFERS

Transfer Buffers
5X transfer buffer(-MeOH)

 

500mL

1L

Tris base

9.37g

18.74g

Glycine

45.0g

90.0g

Blocking Buffer


1X

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


Monoclonal Ab

Polyclonal Ab

1 litre wash buffer plus BSA

1 litre wash buffer plus ( )

0.1% Triton

0.1% Triton

140mM NaCl

560mM NaCl (+ BSA 10g)

10mM KPO4

0.02% SDS (+ 1mL 20% SDS)

0.02% azide

10mM KPO4 (+ 24g NaCl)

1% BSA

0.01% azide

 

1% BSA

Wash Buffer


[1X]

10X (1 litre)

140mM NaCl

80g

10mM KPO4

200mL of 0.5M stock

0.02% azide NaN3

2g

0.1% Triton

10mL

WESTERN AP BUFFER
500 ml 1L 2L

 

500mL

1L

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. *