Your Step by Step Guide to

Total Bacterial Protein and

Membrane Protein Extraction

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Principle and Protocol

The main reagent

Lysate (pH 8.5-9.0): 50 mM Tris-HCl, 2 mM EDTA, 100 mM

NaCl, 0.5% Triton X-100, adjust pH to 8.5-9.0;
Lysozyme; protease inhibitor PMSF; 1% SDS solution; Trizol
lysate; anhydrous ethanol; isopropanol; 0.3M guanidine
hydrochloride;
Hydrophobic protein extract (non-lysing solution): 1% Triton
X-114, 150 mM NaCl, 10 mM Tris-HCl, 1 mM EDTA, and pH
adjusted to 8.0 for use.

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Introduction

This experiment describes several methods for extracting

total bacterial proteins and membrane proteins.

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Major equipment

High-speed centrifuge; Ultrasonic; Dialysis bags; EP tube;

Vortex; Water bath and so on.

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Experimental procedure

1. Extraction of total protein from fresh samples (Simplified

method)
2. Isolation of Total Protein from Trizol Lysate

3. Extraction of hydrophobic membrane proteins from fresh

samples (Triton X-114 detergent method)

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Contact Us

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