Solutions

Published

November 12, 2019

Running buffer (1 liter)

  • 100 ml 10X Tris-Glycine buffer (Final concentration 25 mM Tris, 192 mM Glycine)
  • 10 ml of 10% SDS (final concentration 0.1%)
  • ddH2O to 1000 ml

Do not adjust pH, chill before use.

Transfer buffer (1 liter)

  • 100 ml 10X Tris-Glycine buffer (Final concentration 25 mM Tris, 192 mM Glycine)
  • 100 ml MeOH (Final concentration 10%)
  • ddH2O to 1000 ml

Do not adjust pH, chill before use. The buffer can be re-used up to five times. Make buffer on the day before the experiment to allow to “de-gas”. Disposal: Do NOT pour transfer buffer down the drain, collect in designated bottle and dispose as hazardous waste.

10X TBS (1 liter)

  • 24 g Tris-HCl (formula weight: 157.6 g)
  • 5.6 g Tris base (formula weight: 121.1 g)
  • 88 g NaCl (formula weight: 58.4 g)

Dissolve in 900 mL ddH2O Adjust pH to 7.6 and make up to final volume 1000 ml with ddH2O. The final molar concentrations of the 1X solution are 20 mM Tris and 150 mM NaCl

1X TBS-Tween (1 liter)

  • 100 ml 10X TBS
  • 1 ml Tween-20 (final concentration 0.1%)

To 1000 ml with ddH2O

10X Tris-Glycine buffer (1 liter)

  • 30.3 g Tris (mw 121.14) (Final concentration 250 mM, 25mM in 1X)
  • 144.1 g Glycine (mw 75.07) (Final concentration 1.92 M, 192 mM in 1X)

Dissolve in 800 ml, adjust to final volume 1000 ddH2O. Do not adjust pH, should be around 8.3.

10X Tris-Glycine - Running buffer (1 liter)

  • 30.3 g Tris (mw 121.14) (Final concentration 250 mM, 25mM in 1X)
  • 144.1 g Glycine (mw 75.07) (Final concentration 1.92 M, 192 mM in 1X)
  • 10 g SDS

Dissolve in 800 ml, adjust to final volume 1000 ddH2O. Do not adjust pH, should be around 8.3.

Kopec buffer

Note: The Kopec buffer is suitable for dissolvining the protein pellet after Trizol extraction. See full protocol: Extraction of RNA and protein from muscle samples using Trizol reagent.

Component MW Final concentration For 200 ml
EDTA 372.24 20 mM 1.49 g
NaCl 58.44 140 mM 1.64 g
SDS 5% 10 g
Tris 121.14 10 mM 2.42 g

Adjust pH to 8.0, add phosphatase and protease inhibitors prior to use.