6 Biological buffers recommended for protein purification

forward from: https://www.hopaxfc.com/en/blog/6-biological-buffers-recommended-for-protein-purification

Why purify proteins?

Protein purification is of great research value in the field of biochemistry! Through the purified proteins of interest, researchers can analyze their structures, observe the enzyme activities, study the gene problems as well as develop antibiotics from specific bacterial antibodies, etc.[1]

The buffers recommended for protein purification

pH buffering agents are vital for maintaining stable environmental pH ranges required for protein purification. Under the premise of not interfering with the target protein activity, the types of the target proteins and the purification methods used are the two key factors to consider when choosing proper buffers. Therefore, before determining the buffers that work best to your experiments, you shall decide the methods of purification, the cell samples to be studied and the results you intend to obtain beforehand [2][3]. We hereby introduce you 6 biological buffers commonly used for protein purification, as follows:

1) BIS-TRIS

CAS No.: 6976–37–0
Useful pH range: 5.8–7.2

Uses & Concerns:

◆Used as a running buffer, gel buffer and sample buffer with different types of electrophoresis

◆Used with ACES buffer as a running buffer (BIS-TRIS/ACES) for gel electrophoresis

◆Used for agarose gel electrophoresis

◆Used as a buffer for anion exchange chromatography, NMR spectroscopy and X-ray crystallography.

◆Considered to be an excellent alternate to cacodylic acid, a highly toxic buffer.

◆It interacts with human liver fatty acid-binding proteins (FABPs) and affects the protein dynamics.

◆It forms a strong complex with Pb2+ and Cu2+ and a has a weak interaction with other common metals.

Read more about Hopax BIS-TRIS

2) MOPS

CAS No.: 1132–61–2
Useful pH range: 6.5–7.9

Uses & Concerns:

◆Used in chromatographic methods for protein purification

◆It interacts with the polypeptide backbone of bovine serum albumin (BSA) and affects the protein stability

Read more about Hopax MOPS

3) PIPES

CAS No.: 5625–37–6
Useful pH range: 6.1–7.5

Uses & Concerns:

◆Used in chromatographic methods for protein purification

◆Similar to MES buffer and MOPS buffer, PIPES acts as a non-coordinating buffer for metal ions containing solutions for the lack of ability to form chelate complexes with most metal ions

Read more about Hopax PIPES

4) TES

CAS No.: 7365–44–8
Useful pH range: 6.8–8.2

Uses & Concerns:

◆Used as a buffer for enzyme assays

◆Used for gel filtration chromatography and affinity chromatography

Read more about Hopax TES

5) TRIS

CAS No.: 77–86–1
Useful pH range: 7.2–9.0

Uses & Concerns:

◆Used for anion exchange chromatography

◆Used in buffer solutions (TAE or TBE) for gel electrophoresis

◆Used in the preparation of Laemmli buffer for denaturing and loading of protein samples in SDS-PAGE

◆Not suitable for use with the Bicinchoninic acid (BCA) assay

◆It may interfere with the Bradford dye-binding method of protein assay

Read more about Hopax TRIS

6) TRIS-HCL

CAS No.: 1185–53–1
Useful pH range: 7.2–9.0

Uses & Concerns:

◆Used for hydrophobic interaction chromatography (HIC) as a ingredient of the separation and dialysis solutions

◆Used for sample loading of ion exchange chromatography

◆Used in the substrate solution of affinity chromatography

◆Used for SDS-PAGE as an ingredient of the elution buffer

Read more about Hopax TRIS-HCL

As a professional biological buffer manufacturer, HOPAX produces more than 30 high-quality buffers, offers a variety of packaging options as well as friendly and comprehensive services. Should you have any requirements related to HOPAX buffers, please feel free to contact us!

References:

1 Studying Proteins and Protein Purification

2 Protein Lysate TLC: Pro-Tips to Keep Your Protein Extracts in (Experimentally) Perfect Shape.

3 How to choose the perfect buffer to get a pure, stabilised, functional protein

4 Purification and biochemical characterization of a novel thermostable serine alkaline protease from Aeribacillus pallidus C10: a potential additive for detergents Vildan Yildirim, a Mustafa Ozkan Baltaci, a Ilknur Ozgencli, b Melda Sisecioglu, a Ahmet Adiguzel, a and Gulsah Adiguzel c

5 Native RNA purification by gel filtration chromatography.

6 Protein Stability, M. Michael Gromiha, in Protein Bioinformatics, 2010