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SDS PAGE: REAGENTS

Gel-Making Solutions:
1. 30% acrylamide/0.8% N,N-methyl-bis-acrylamide solution (4C storage in dark bottle,
1month) : 150.00 mL
Acrylamide
N,N-bis-methylene-acrylamide
Di H2O

43.80 g
1.20 g
Bring up to 150mL

2. Buffer A : 1.5 M Tris-HCl pH 8.8 (4C storage): 150.00 mL

Tris-Base

27.23 g
o Add 80mL Di H2O then adjust pH with HCl
o Bring up to 150.00 mL

3. Buffer B : 0.5 M Tris-HCl, pH 6.8: 100.00 mL

Tris-Base

6.00 g
o Add 80mL Di H2O then adjust pH with HCl
o Bring up to 100mL
4. 10% SDS (w/v): 100.00 mL
SDS

10.00 g
o Add 90mL of Di H2O and gently stir to dissolve
o Bring up to 100.00 mL
5. 10% APS (make fresh weekly, store at 4C): 1.00 mL
Ammonium persulfate
Di H2O

0.10 g
1.00 mL

6. Water-Saturated n-Butanol: 5.00 mL

n-Butanol
Di H2O

50.00 mL
5.00 mL

SDS PAGE: REAGENTS

Gel-Running Solutions:
7. 10X Running Buffer (pH 8.3, no adjustments is required, keep @ RT): 4.00 L
Tris-Base
Glycine
SDS
Di H2O

121.10 g
576.00 g
200.00 mL of a 20% SDS solution
---OR--40.00 g
Bring up to 4.00 L

8. 10X Transfer Buffer (Western Blotting): 4.00 L

Tris-Base
Glycine
Di H2O

121.10 g
576.00 g
Bring up to 4.00 L

9. 20X TBS Buffer pH 7.6 (Western Blotting): 4.00 L

Tris-Base
NaCl

193.60 g
640.00 g
o Add 3.20 L Di H2O then adjust pH with HCl
o Bring up to 4.00 L
10. 20X TBST Buffer (Western Blotting): 100.00 mL
Tween-20
20X TBS pH 7.6

2.00 mL
Bring up to 100.00 mL

SDS PAGE: REAGENTS

Other Solutions
11. 2X Sample Loading Buffer (non-reducing): 100.00 mL
1 M Tris, pH 7
20% SDS
Glycerol
Bromophenol Blue
Di H2O

5.00 mL
25.00 mL
20.00 mL
2.00 mg
Bring up to 100.00 mL

12. 2X Sample Loading Buffer (reducing); 1.00 mL

2X non-reducing sample loading buffer
-mercaptoethanol

950.00 uL
50.00 uL

13. Stain/De-stain Solution: 4.00 L

Absolute Ethanol
Glacial Acetic Acid
Di H2O

200.00 mL
300.00 mL
Bring up to 4.00 L

14. Fixing Solution: 1.00 L

Absolute Ethanol
Glacial Acetic Acid
Di H2O

600.00 mL
300.00 mL
Bring up to 4.00 L

15. Coomassie Blue Stock Solution (G-250): 100.00 mL

Coomassie Brilliant Blue (G-250)
Di H2O

250.00 mg
100.00 mL