Tris-Glycine Transfer Buffer

Author

Affiliations

Pranav Kumar Mishra

Dept of Surgery, Rush University

Dept of Orthopedic Surgery, Rush University

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