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For most standard sizes, see our general Western Blot protocol.
Prepare solutions, reagents and the gels according to the recipes below.
10 x running buffer |
| Dissolve compoments in 3.5 L of water initially, then make up to 5 L Dilute 1:10 in to 1X when ready for use |
1X transfer buffer |
| Dissolve compoments in 3.5 L of water initially, then make up to 10 L Add 10 % methanol to transfer buffer just before use |
Blocking buffer |
| |
10X TBS |
| Dissolve in deionized water, adjust pH to 7.4 using HCl, then make up to 22.5 L with deionized water. |
1X TBST |
| Add 2.25 L 10X TBS and 22.5 L Tween-20, make up to 22.5 L with deionized water |
Separation gel |
| |
Stacking gel |
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The time and voltage may require optimization. We recommend following the manufacturer’s instructions. A reducing gel should be used unless non-reducing conditions are recommended on the antibody datasheet.