beautypg.com

Bio-Rad Trans-Blot® Plus Cell User Manual

Page 22

background image

Dunn carbonate buffer

In some cases, this buffer may produce higher efficiency transfers and improve

the ability of antibodies to recognize and bind to proteins.

10 mM NaHCO

3

, 3 mM Na

2

CO

3

, 20% methanol, pH 9.9

a) Using dry reagents:

0.84 g NaHCO

3

0.318 g Na

2

CO

3

(anhydrous)

500 ml deionized water (dd H

2

O)

200 ml methanol

add dd H

2

O to I liter

2. Buffers for native gels

Towbin Buffer

25 mM Tris, 192 mM glycine, pH 8.3

a) Using 10X Tris /glycine buffer (catalog #161-0734 for 1L bottles or
catalog # 161-0757 for 5L cube)

100 ml of 10X Tris /glycine buffer

900 ml of deionized water (dd H

2

O)

b) Using dry reagents:

3.03 g Tris

14.4 g glycine

600 ml deionized water (dd H

2

O)

add dd H

2

O to 1 liter

3. Buffers for nucleic acid transfers

TBE (Tris-Borate EDTA)

89 mM Tris borate, 2 mM EDTA pH 8.3

a) Using 10X TBE buffer (catalog #161-0733 for 1L bottles or
catalog # 161-0770 for 5L cube)

100 ml of 10X TBE buffer

900 ml of deionized water (dd H

2

O)

b) Using dry reagents (10X stock)

108 g Tris base

55 g boric acid

40 ml 0.5 M EDTA, pH 8.0

Add 100 ml of the 10X stock to 900 ml deionized water (dd H

2

O)

to make a 1X working solution.

20