Physical and chemical properties of Tris buffer

Tris buffer is a commonly used biological buffer with the following physicochemical properties and composition:

Composition: Tris buffer is mainly composed of trimethylolaminomethane (TriS) and hydrochloric acid (HCl) or sodium hydroxide (NaOH). Tris is an organic compound that has an amino group and three hydroxyl groups that can act as a component of buffers.

Physical and chemical properties: 1) pH range: The pH range of TRIS buffer is usually between to, and the pH value can be adjusted by adding hydrochloric acid or sodium hydroxide.

2) Buffer capacity: TRIS buffer has a large buffer capacity and is able to maintain a stable pH value over a wide pH range.

3) Ionic strength: Tris buffer has a low ionic strength and does not cause significant interference to biomolecules.

4) Solubility: TRIS buffer is easily soluble in water and can dissolve biomolecules well.

5) Stability: TRIS buffer can be stored for a long time at room temperature, and it is not easy to decompose or deteriorate.

Purpose: TRIS buffer is mainly used to maintain pH stability in biochemical experiments to ensure the accuracy of experimental results. For example, TRIS buffers can be used as components of eluents, lytics, or reactions in experiments such as protein purification, DNA and RNA extraction.

Overall, TRIS buffer is a stable and reliable biological buffer with a wide range of applications.

Preparation method of Tris-HCl buffer:

After mixing 50ml of tris-hydroxymethyl aminomethane (TriS) solution with x ml of hydrochloric acid, dilute to 100ml with water to successfully configure.

Tris(hydroxymethyl)aminomethane (Tris) is an organic compound with the molecular formula (Hoch2)3CNH2.

Tris is a weak base, and at room temperature (25, its PKA is; According to the buffer theory, the effective buffer range of TRIS buffer is between to.

The aqueous solution of Tris base is around pH, and hydrochloric acid is generally added to adjust the pH value to the desired value, and the buffer of this pH value can be obtained. At the same time, however, attention should be paid to the effect of temperature on the PKA of Tris.

Uses:

TRIS buffers are not only widely used as solvents for nucleic acids and proteins, but also have many important uses. Tris was used for protein crystal growth at different pH conditions. The low ionic strength characteristic of TRIS buffer can be used in nematodes (C

elegans) formation of intermediate fibers of lamin. Tris is also one of the main components of protein electrophoresis buffers, which form a buffer system with glycine in the electrophoresis buffer to stabilize the pH during electrophoresis.

Tris HCl buffer is formulated as follows:

1. 1 M Tris-HCl (pH, prepare 1000ml

Preparation: Weigh Tris in a 1000 ml beaker. Add about 800ml of deionized water, stir and stir well to dissolve.

Add the amount of concentrated HCl in the table below to adjust the pH value of the acorn. Bring the solution to 1000 ml. After autoclaving, store at room temperature.

Note: The pH of the solution should be cooled to room temperature before setting, as the pH of the Tris solution varies greatly with temperature, and the pH of the solution decreases by about one unit for every 1 increase in temperature.

2. Tris-HCl (pH prepared 1000ml

Preparation method such as staring: weigh tris and place it in a 1000ml beaker. Add about 800ml of deionized water and stir well to dissolve.

Adjust the pH to with concentrated HCl. Bring the solution to 1000 ml. After autoclaving, store at room temperature.

Note: The pH of the solution should be cooled to room temperature before setting, as the pH of the Tris solution varies greatly with temperature, and the pH of the solution decreases by about one unit for every 1 increase in temperature.

3. TE is Tris-EDTA Buffer (10mm Tris, 1mm EDTA, preparation method: take the following solution and place it in a 1000ml beaker. 1 M Tris-HCl Buffer (,,500 mM EDTA) (Add about 800 ml of deionized water to the beaker and mix evenly.

After the solution was fixed to 1000ml, it was sterilized by high temperature and autoclaving. Store at room temperature.

Tris-HCl is a hydrochloride made on the basis of Tris.

Tris-HCl buffer is prepared as follows:

Using a graduated cylinder or volumetric flask, measure the water you need. According to the needs of the experiment, determine the volume of the required buffer and add the appropriate amount of water to the container. Be sure to use purified or deionized water to avoid impurities affecting the results.

Add the Tris base to the container. Calculate the mass or volume of the required TRIS base at the desired concentration for the experiment and slowly add it to the stirred water. Note that Tris base has limited solubility, so it is recommended to stir slowly at room temperature and wait for it to dissolve completely.

Adjust the pH level. Hydrochloric acid (HCl) is added dropwise to the buffer while monitoring the pH of the solution using a pH meter. According to the experimental requirements, adjust the pH value to the desired range.

Care should be taken when adjusting the pH to avoid adding too much hydrochloric acid and causing the solution to become too acidic.

Stir and store. Use a magnetic stirrer or stir bar to stir the solution well to ensure that the components are evenly mixed. Once the preparation is complete, the buffer can be transferred to an appropriate container and stored as needed.

The advantages of Tris-HCl buffer are:

Because the alkalinity of TRIS base is strong, it is possible to prepare buffers with a wide range of pH values from acidic to alkaline with only this buffer system;

It has little interference with biochemical processes and does not precipitate with calcium, magnesium ions and heavy metal ions.

Its disadvantages are: the pH value of the buffer is greatly affected by the concentration of the solution, the buffer is diluted ten times, and the change of the pH value is greater;

The temperature effect is large, and the temperature change has a great influence on the pH value of the buffer, i.e.:

For example, the pH of the buffer at 4 and the pH at 37 must be prepared at the use temperature, and the Tris-HCl buffer prepared at room temperature cannot be used for 0 4.

It is easy to absorb CO2 in the air, so the prepared buffer should be tightly sealed.

This buffer interferes with some pH electrodes, so use an electrode that is compatible with TRIS solutions.