Tris is a widely used organic reagent with an effective buffer range of pH 7.0 to 9.2. It and its derivatives are widely used in the preparation of buffer solutions in biochemistry and molecular biology experiments, and can be used in the production of drugs, organic synthesis, biological buffers and so on.
In biochemical experiments, many protein and nucleic acid related experiments need Tris as a biological buffer. It even has a tendency to surpass phosphate buffer. This article will briefly describe its derivative buffer.
TBS is Tris buffer salt water, which is Tris HCl buffer salt solution. Because Tris base has strong alkalinity, it can only be used to prepare buffer solution with a wide range of pH from acidic to alkaline, which has little interference to biochemical process and does not precipitate with calcium, magnesium ions and heavy metal ions. However, the pH value of the buffer solution is greatly affected by the concentration of the solution, the temperature effect is also great, and it is easy to absorb CO2 in the air, so the prepared buffer solution should be tightly sealed, and this buffer solution has a certain interference effect on some pH electrodes, so the electrode compatible with tris solution should be used.
Tbst contains Tris HCl, NaCl and Tween20, which is a common membrane washing buffer in Western blotting. The aim is to wash away non-specific proteins and maintain a close natural environment. Tween is an ionic surfactant, which can accelerate the separation of nonspecific antibody binding. Because the membrane used in Western blotting can bind to protein, the antibody can bind to the membrane nonspecificly during incubation. In order to prevent the film background from being too high due to later exposure, TBS mixed with Tween 20 can wash away the non-specific binding antibody after incubating the antibody to increase the specificity.
Te buffer is made by adding EDTA into Tris hydrochloric acid buffer. Te buffer is weakly alkaline and has protective effect on DNA base. Therefore, DNA is stable in te, and it is not easy to damage its integrity or produce ring opening and breakage. Te buffer is used for DNA stability and storage.
The most widely used buffer system is TAS / acetate / EDTA. It is characterized by the fact that the super helix electrophoresis is more in line with the actual molecular weight, and the mobility of double stranded linear DNA in it is also faster. When electrophoresis fragments larger than 13KB, Tae buffer will achieve better separation effect. In addition, it is easy to use Tae buffer system for electrophoresis when recovering DNA fragments. However, the disadvantage of TAE is that the buffer capacity is small. Unless there is a circulation device to exchange the buffer solution between the two poles, long-time electrophoresis is not optional.
Acrylamide experiments and agarose gel electrophoresis were used to convert the acid solution of pH to boric acid, and the "TBE buffer" (Tris/Borate/EDTA) was obtained. Tbe buffer is a common buffer in molecular experiments, which is composed of Tris, boric acid and EDTA, so it is called tbe buffer for short. The buffer is often used in the experiment of polyelectrophoresis. Tbe buffer is a colorless clear liquid, which can be stored at room temperature. The pH value of tbe buffer is 8.2 ~ 8.4 at 25 ℃. It should be noted that tbe buffer can not contain the activity of DNA hydrolase and RNA hydrolase.
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