Transmembrane Buffer (10×)

Introduction

Tris-Glycine transmembrane buffer is used in the wet and semi-dry test of Western blotting experiments. The protein can be transferred from the gel to the PVDF membrane or the NC membrane. This product is supplied with 10 × concentrate for easy transportation (without methanol).

Components

Instructions

This product is a 10× concentrate, dilute with Deionized water and methanol to 1× working solution before use. 

For example：take 100 mL Transmembrane Buffer (10×), 200 mL methanol,dilute with Deionized water to 1 L.

In general, 20% methanol is recommended and can be adjusted according to specific situations. If high molecular weight protein transfer is not easy, please adjust the concentration of methanol. For proteins with a molecular weight larger than 200 kDa, the methanol concentration can be reduced from 20% to 5%, and the transmembrane time can be appropriately increased to 3 h.

Self-Prepared Reagent

Methanol

1 × Transmembrane Buffer Components

25 mM Tris-base, 192 mM Glycine, 20% (v/v) Methanol, pH8.3.

Storage

Store at 2~8°C for 12 months.

Cautions

1.This product does not provide methanol, but it is required.

2.This product is a 10 × concentrate solution. Once it is formulated as a 1 × working solution and methanol is added, please use it within one week.

3.This product does not contain SDS. For proteins with a molecular weight larger than 120 kDa or more hydrophobic, (0.025-0.1%) SDS can be added to prevent protein from accumulating in the gel.

4.This product is a high concentrate. When the temperature is low, crystals may be precipitated. It can be heated to dissolve. Dilute after it is completely dissolved.

5.Keep the product sealed to prevent from pollution.

6.For your safety and health, please wear the lab coat and disposable gloves before the experiments.