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Preparation of Tris-HCL buffer and its advantages and disadvantages

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Preparation of Tris-HCL buffer and its advantages and disadvantages

Classification:
News
Author:
2020/06/22 16:00

Tris-HCL Chinese name tris (hydroxymethyl) aminomethane hydrochloride, is a chemical substance, its chemical formula is C4H11NO3.HCl. It is prepared by adding Tris to hydrochloric acid. The commonly configured 1M Tris-HCl 6.8 and 1.5M Tris-HCl 8.8 are the most commonly used reagents for SDS-PAGE. The specific configuration method is as follows:

Tris-HCL powder and packaging

1. Tris-HCl (pH7.4, 7.6, 8.0)

Component concentration: 1 M Tris-HCl

Preparation volume: 1 L

Preparation method:

1. Weigh 121.1 g Tris into a 1 L beaker

2. Add about 800 ml of deionized water, stir well to dissolve

3. Add concentrated hydrochloric acid to adjust the required pH

4. Bring the volume to 1 L

5. After autoclaving, store at room temperature

2. 1.5 M Tris-HCl (pH8.8)

Component concentration: 1.5 M Tris-HCl

Preparation volume: 1 L

Preparation method:

1. Weigh 181.7 g Tris into a 1 L beaker

2. Add about 800 ml of deionized water, stir well to dissolve

3. Adjust pH to 8.8 with concentrated hydrochloric acid

4. Bring the volume to 1 L

5. After autoclaving, store at room temperature

The advantages of Tris-HCl buffer:

1. Because Tris base is more alkaline, you can use this buffer system to prepare a buffer solution with a wide range of pH values ​​from acidic to alkaline;

2. Little interference to biochemical process, no precipitation with calcium, magnesium ions and heavy metal ions.

Disadvantages of Tris-HCl buffer:

1. The pH value of the buffer solution is greatly affected by the concentration of the solution. The buffer solution is diluted ten times, and the change in pH value is greater than 0.1;

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

△pKa/℃=0.031, for example: the buffer pH at 4℃=8.4, then the pH at 37℃=7.4, so it must be prepared at the use temperature, and the Tris-HCl buffer prepared at room temperature cannot be used At 0 ℃ ~ 4 ℃.

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

Since its establishment in 2005, Desheng has researched biological buffers, blood collection tube additives, color reagents, and chemiluminescent reagents. It has rich production experience and can provide customers with high-quality raw materials for products. In addition to Tris-HCL, there are TRIS, HEPES, CAPS, PIPES, etc. Biological buffers can also be configured with different concentrations of buffers for customer needs