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The use and preparation method of biological buffering agent PIPES powder

2024-01-04
The use and preparation method of biological buffering agent PIPES powder

Biological buffer PIPES powder is a chemical reagent widely used in biological experiments, and its full name is piperazine-1,4-diethanesulfonic acid. As a biological buffer, PIPES solution plays an important role in maintaining the stability of the intracellular environment and protecting cells from pH fluctuations. This article will provide a detailed introduction to the properties, uses, and preparation methods of the biological buffering agent PIPES powder.

 

PIPES is a white powdered chemical reagent with the chemical formula C8H12N2O4S and a molecular weight of 248.26. It is a zwitterionic buffering agent with stable pH buffering ability, capable of maintaining a certain pH value in acidic or alkaline environments. In addition, PIPES also has high ion strength and low permeability, making it widely used in biological experiments such as cell culture, electrophoresis, and immunoblotting.

 

The application of biological buffer PIPES in biological experiments

 

PIPES powder, as a biological buffer, has been widely used in many biological experiments. The following are several typical application areas:

 

1. Cell culture: In cell culture experiments, PIPES can serve as a pH buffer to maintain the pH stability of the culture medium and ensure normal cell growth and division. It can serve as an ion carrier to facilitate ion exchange inside and outside the cell, affecting the physiological functions of the cell.

 

2. Electrophoresis: In electrophoresis experiments, PIPES powder can be used as a molecular sieve and ion exchanger to improve the resolution and effectiveness of the experiment. For example, in protein electrophoresis experiments, PIPES can bind to proteins, slowing down their movement in an electric field, thereby improving the resolution of the experiment

 

3. Immunoblotting: In the immunoblotting experiment, PIPES can be used as the main component of the transmembrane buffer to help transfer proteins from the gel to the membrane. Meanwhile, PIPES can also serve as an ion carrier to improve the sensitivity and specificity of experiments.

 

4. Purification of microtubule proteins: In microtubule protein purification experiments, PIPES can serve as the main component of the buffer to assist in the extraction and purification of microtubule proteins from tissues. It can also inhibit the polymerization reaction of microtubule proteins and improve purification efficiency.

 

5. Peroxisome localization: In peroxisome localization experiments, PIPES can serve as the main component of the buffer to assist in the ultrastructural localization of peroxisomes in cells. It can also protect cells from damage by peroxisomes.

 

In addition to the aforementioned application areas, PIPES can also be applied in biological experiments such as the separation and purification of nucleic acids and proteins, protein crystallization, tissue culture, and drug screening. In summary, as an important biological buffer, it plays an indispensable role in many biological experiments.

 

Preparation method of biological buffer PIPES

 

1. Prepare the required reagents and equipment

 

PIPES powder, ultrapure water, glass bottle, weighing paper, electronic balance, magnetic stirrer or vortex oscillator.

 

2. Weighing PIPES powder

 

Accurately weigh PIPES powder using an electronic balance based on the required concentration and volume. For example, if 100 mL of 0.25 mol/L PIPES solution needs to be prepared, 25.7g of PIPES powder needs to be weighed.

 

3. Dissolve PIPES powder

 

Add the weighed PIPES powder into a glass bottle, add an appropriate amount of ultrapure water, and then stir with a magnetic stirrer or vortex oscillator until dissolved. Note that sufficient ultrapure water should be used to ensure complete dissolution.

 

4. Filtering and sterilization

 

Filter and sterilize the prepared PIPES solution to remove impurities and bacteria from the solution. Disposable syringes or filters can be used for filtration.

 

5. Storage and Use

 

Store the filtered PIPES solution in a clean glass bottle and store it in a refrigerator at 4 ℃. When using, an appropriate amount of PIPES solution should be added according to experimental requirements.

 

Precautions for the use of biological buffer PIPES

 

1. PIPES powder has hygroscopicity, so it should be stored in a dry environment to avoid moisture;

 

2. During the configuration process, ultrapure water or deionized water should be used to ensure the purity and quality of the solution;

 

3. The prepared PIPES solution should be a colorless and transparent liquid. If turbidity or precipitation occurs, it may be due to incomplete dissolution of the powder or the presence of impurities, and it should be reconfigured;

 

4. The prepared solution should be used up within the specified time to avoid prolonged storage that may affect the quality and effectiveness of the solution.

 

Desheng is a manufacturer of PIPES powder with high purity and stable quality, which can be provided in large quantities. We can provide over 30 different types of buffer products, and if you need other biological buffer, you can also purchase them in one stop. Feel free to contact us anytime if you need anything!