The above mentioned recipe is perfect for 10ml. This buffer can be created in large quantities and utilized as a wash buffer, or allotted and frozen for future use. Immediately before use add protease inhibitors in the manufacturers suggested concentration, and phosphatase inhibitors if needed.
Laemmli's Buffer, 4x
- 2.4 ml 1 M Tris pH 6.8 (Just like upper gel buffer)
- .8 g SDS stock
- 4 ml 100% glycerol
- .01% bromophenol blue. Final Concentration is .02%
- 2.8 ml ddH2 O
Before use add 1/10 th amount of -mercaptoethanol
Laemmli's Buffer, 6x
- 1.2g SDS (sodium dodecyl sulfate)
- .01% bromophenol blue
- 4.7ml glycerol
- 1.2ml Tris .5M pH6.8
- 2.1ml ddH2 O
Before use add 1/8 th amount of -mercaptoethanol
4x Lower Gel Buffer
pH to eight.8 don't overshoot. Use glass pipettes to pH.
4x Upper Gel Buffer
pH to six.8, don't overshoot. Use glass pipettes.
10x Running Buffer
Dissolve in 1L dH2O. Dilute 1:10 in ddH2O to be used as running buffer
10X Transfer Buffer (Towbin Buffer)
Bring up to 1L in ddH2O.
1X Transfer Buffer (Towbin Buffer)
10x Tbsp .
Dilute to 5L in dH2O, pH to 7.5 with saturated HCl.