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Western Blot Gels

Western Blot Gels[]

Brief Description

Gel percentages and instructions for pouring gels

Protocol Details

Western Blot Gel Set-up

SDS-PAGE GELS

 

Separating Gel

%

5

6

7

7.5

8

9

10

12

13

15

16.5

30% acrylamide

1.67

2

2.33

2.5

2.67

3

3.33

4

4.33

5

5.5

separating buffer

2.5

2.5

2.5

2.5

2.5

2.5

2.5

2.5

2.5

2.5

2.5

ddi H2O

5.83

5.5

5.17

5

4.83

4.5

4.17

3.5

3.17

2.5

2

10% APS 50uL

TEMED 12uL

 

Stacking Gel

30% acrylamide .67mL
stacking buffer 1.25mL
ddi H20 3mL

10% APS 30uL

TEMED 7uL

 

Note: above volumes are for two gels

-Add 10% APS and TEMED just before pouring gel

-Pour separating gel, ~5mL /gel and add a small volume of t-butanol or 2-propanol and let stand ~15 minutes until gel polymerizes

-After gel polymerizes, pour off t-butano. Remove excess with P200 pepette and paper towel

-Pour stacking gel and let stand until polymerized

 

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