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Buffering agents containing structure of Tris (77-86-1)

2017-10-23 来源:亚科官网
23 October 2017
  Bis-Tris, Tricine, TES, TAPS are the buffers commonly used in biochemical experiments and molecular biology experiments, and their molecular structures contain that of Tris. So what are the differences between these buffers and Tris? What are the differences between them? This paper will summarize these types of buffers.
Tris
Tris
(1) commonly used in gel electrophoresis buffer, such as TAE or TBE;
(2) commonly used in the preparation of Laemmli buffer;
(3) prepare running and loading buffer with glycine and SDS;
(4) used as binding buffer and eluent in anion exchange chromatography;
(5) used as a buffer for RNA hybridization and DNA cleavage.
  In the use of Tris, please note: the temperature and concentration has great impact on the pH value, so the preparation process should consider the environment; primary amino group on Tris, to a certain extent, has reaction with a variety of moleculars, including RNA Inhibitors, aldehydes, common metals such as Cr3+, Fe3+, Ni2+, Co2+ and Cu2+, enzymes, and DNA, in addition, Tris is not suitable for use with the Bicinchoninic acid (BCA) assay.
Bis-Tris
Bis-Tris
  Bis-Tris, zwitterionic buffer, pKa=6.46 at 25°C and Ph range 7.2~7.2. Its application areas are as follows:
(1) sample buffer, gel buffer and running buffer for various types of electrophoresis;
(2) buffer in anion exchange chromatography;
(3) X-ray crystallography studies for the crystallization process of haloalkane dehalogenase;
(4) low conductivity and high sensitivity, is an effective buffer for NMR spectroscopy;
(5) interacts with human hepatic fatty acid binding protein (FAB) and affects protein kinetics.
  Bis-Tris can form strong complexes with Cu2+ and Pb2+, form weak complexes with many common metals, so its complexing constants should be considered when used in metal ion-containing solutions. In addition, Bis-Tris buffer solutions can be used in systems with pressure changes. It is also not suitable for use with the Bicinchoninic acid (BCA) assay.
Tricine
  Tricine can combine hydroxyl radicals to form formaldehyde, or form trinitroxygen free radicals, which should be taken into account in the presence of enzymes with oxidase activity. Tricine forms strong complexes with many common metals, such as Mg2+, Ca2+, Ni2+, Co2+, Zn2+ and Cu2+, so the stability constants and concentrations should be taken into account when selecting the buffer. Tricine is also not suitable for use with the Bicinchoninic acid (BCA) assay.
TES
TES
  Zwitterion buffer, pKa (25°C) is 7.40, buffer range 6.8~8.2. Because the buffer range is just in the physiological conditions, many biologists preferred to use the buffer. The common experiment is as follows:
(1) cell wash buffer;
(2) solution for precipitation and extraction of DNA;
(3) for the purification of microsomes from porcine kidney by gel filtration and affinity chromatography;
(4) for enzyme assay buffer.
  Like most neutral amine-based buffers, TES can form complexes with DNA; its ability to limit enzyme kinetics is stronger than HEPES, but weaker than Tris; TES forms strong complexes with common metals such as Cr3+ and Fe3+, weak complexes with Co2+, Ni2+, Cu2+ and Zn2+. TES is suitable for use with the Bicinchoninic acid (BCA) assay.
TAPS
TAPS
(1) for the chromatographic separation of dyes;
(2) for the separation and purification of glutathione S-transferase by ion exchange chromatography;
(3) determining the aminopeptidase activity as an enzyme assay buffer;
(4) preferred medium buffer for flagellate experiments.
  TAPS forms strong complexes with Cr3+, Fe3+ and Cu2+, forms weak complexes with other common metals, so the stability constants and concentrations should be taken into account when selecting the buffer.
  In specific use, buffers containing the Tris structure will form a strong or weak complex with a variety of metal ions, so the stability constant should be taken into account. In addition, the buffer range and the appropriate type of experiment also should be considered in order to obtain the best experimental results.
Related links: Tris
                       Bis-Tris
                       Tricine
                       TES
                       TAPS
Edited by Suzhou Yacoo Science Co., Ltd.