Transfer Buffer for Western Blot

-
10X Transfer Buffer
30.3g Tris
144.1g Glycine
water to 1L

1X from stock
700 mls water
100 mls 10x
200mls methanol
makes 1L

Comments

Post a Comment

Popular posts from this blog

Prepare 6X Laemmli buffer

-
Laemmli is a sample buffer to use in western blot. 6X sample buffer is added to each protein sample and is boiled or heated for 5-10 minutes. Recipe to prepare 10 ml: - 1.2gr SDS (sodium dodecyl sulfate) - 6mg bromophenol blue - 4.7ml glycerol - 1.2ml Tris 0.5M pH6.8 - 2.1ml dwater warm it a little bit and shake it till everything is dissolved. It takes time. - add 0.93gr DTT. Wait until it is completely dissolved. Aliquote and keep frozen at-20ºC.
Read more

0.1 M Triethanolamine

-
0.1 M TEA: TEA = Triethanolamine You will dilute TEA 1:10 in Millipore water: Pour 50 mL TEA into a 50 mL conical tube (TEA is very viscuous, so best to measure it in something that is disposable). Pour TEA from conical tube into 450 mL Millipore water with stirring. pH to 8.0
Read more

100% Steinberg’s Solution

-
100% Steinberg’s Solution (AKA 5xMR) (makes 2 liters) 40 ml NaCl stock solution (17% w/v NaCl) 20 ml KCl stock solution (0.5% w/v KCl) 20 ml CaCl2 stock solution (0.5% w/v CaCl2) 20 ml MgSO4 stock solution (1.025% w/v MgSO4) 1 ml gentamycin stock solution (4% w/v gentamycin) 2.4 g HEPES 1899 ml ddH2O Adjust pH to 7.4 - 7.6 and store in refrigerator. Usually, you will make 2-4 liters at a time. To make the required stock solutions,
Read more