I don't know, I am not a biochemist. It is how most people go about it.sotellme said:
sotellme said:i just don't know how can 25 mM Tris-base added to 25 mM phosphoric acid and still we get 25mM buffer. won't the concentration of each of the solutions change?
Tris-phosphate buffer is a commonly used buffer solution in biochemistry and molecular biology experiments. It is made up of two components, Tris (tris(hydroxymethyl)aminomethane) and phosphate, which help maintain a stable pH for reactions and prevent drastic changes in pH due to added acids or bases.
A pH of 7.8 is considered optimal for many biological applications as it is close to the physiological pH of most living organisms. This pH range allows for optimal enzyme activity and stability, which is crucial for many biochemical reactions.
To make a 25 mM Tris-phosphate buffer, you will need to mix Tris base and monobasic sodium phosphate in the desired proportions. The exact amounts will depend on the final volume of buffer needed and the concentration of the stock solutions. It is recommended to use a buffer calculator or consult a buffer preparation guide for specific instructions.
Yes, the pH of the buffer can be adjusted by adding either an acid or a base. To increase the pH, add a basic solution such as sodium hydroxide or potassium hydroxide. To decrease the pH, add an acidic solution such as hydrochloric acid or acetic acid. It is important to use a pH meter or indicator to ensure the desired pH is reached.
Tris-phosphate buffer should be stored in a tightly sealed container at room temperature. It is recommended to make fresh buffer as needed for each experiment, but if stored properly, it can be used for up to a month. However, it is important to check the pH before using the buffer as it may change over time.