Tris-Glycine Transfer Buffer

Author
Affiliation

Pranav Mishra

Rush University

Published

Friday, June 9, 2023

Overview

Tris-Gycine Transfer Buffer has a pH of 8.3. Protocol adapted from Abcam1

Uses

Transfer buffer used in Western Blots

Components

  • Tris Base
  • Glycine
  • \(HCl\)
  • 100% Methanol
  • \(diH_2O\)

Protocol

10x Stock of Transfer Buffer

  1. Add 700 mL of \(diH_2O\) to a beaker
  2. Dissolve 30.25 g of Tris Base and 142.63 g of Glycine in the water
  3. Adjust pH with \(HCl\) to 8.3
  4. Add 300mL of \(diH_2O\) (total vol 1000 mL)

1x Transfer Buffer

  1. Add 700 mL of \(diH_2O\) to a beaker
  2. Add 100 mL of 10x Stock Transfer Buffer
  3. Adjust pH with \(HCl\) to 8.3
  4. Add 200mL of 100% Methanol (total vol 1000 mL)

1x concentrations: - Tris 25 mM - Glycine 190 mM

Notes

Concentrations of Tris and Glycine differ from ThermoFicher’s protocols2, which recommend a 12.5 mM of Tris and 96 mM Glycine.

Component Abcam (mM) Thermo (mM)
Tris base 25 12.5
Glycine 190 96

References

  1. Transfer and staining of proteins in western blot. Abcam. https://www.abcam.com/protocols/transfer-and-staining-of-proteins-in-western-blot
  2. Western Blot Protocols and Recipes. ThermoFicher Scientific. https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-biology-learning-center/protein-gel-electrophoresis-information/western-blot-protocols.html

Supplemental

Key resources table

REAGENT OR RESOURCE SOURCE IDENTIFIER
Chemicals
Tris Base Ficher Cat# BP154-1
Glycine Sigma Cat# G-7126
100% Methanol Ficher Cat# A412-4
HCl Lab Stock N/A

Citation

BibTeX citation:
@online{mishra2023,
  author = {Mishra, Pranav},
  title = {Tris-Glycine {Transfer} {Buffer}},
  date = {2023-06-09},
  url = {https://github.com/pranavmishra90/research-reference/protocols/recipes/buffers/tris-glycine-transfer-buffer.html},
  langid = {en-US}
}
For attribution, please cite this work as:
Mishra, Pranav. 2023. “Tris-Glycine Transfer Buffer.” June 9, 2023. https://github.com/pranavmishra90/research-reference/protocols/recipes/buffers/tris-glycine-transfer-buffer.html.