Preparation method of Bicine, HEPES and EPPS buffer

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Biological buffer is one of the most widely used auxiliary reagents in biochemical experimental research. Among them, the more typical buffers are dihydroxyethylglycine Bicine, hydroxyethylpiperazine ethanesulfonic acid HEPES, and hydroxyethylpiperazine probesulfone. Acid EPPS (or HEPPS) and so on.


Biological buffers like these do not directly participate in the reaction or affect the detection-related reactions in the reaction system. They are used to adjust the pH value of the reaction environment, but their effect is indispensable. Most biochemical reactions are originally carried out in organisms, and usually need to be catalyzed and adjusted by enzymes, and the activity of enzymes needs to be displayed at a suitable pH. Each organism has its own system to adjust the pH value. For example, the human body can adjust the pH through the level of CO2 and HCO3-. Then, there is no need to use buffers such as Bicine, HEPES, EPPS, etc. to maintain a proper pH during in vitro experiments.



Preparation of Bicine, HEPES and EPPS buffers

The preparation method of Bicine buffer:

(1) With 0.1M solution (A): Bicine16.317g/ deionized water 1,000ml

(2) With 0.1M NaOH solution (B): NaOH4g/ deionized water 1,000ml





pH 10.41,000ml(A)+800ml(B)(A):(B)=5:4


The preparation method of HEPES buffer:

(1) With 0.1M solution (A): HEPPS25.233g/ deionized water 1,000ml

(2) With 0.1M NaOH solution (B): NaOH4g/ deionized water 1,000ml

(3) Similar to the preparation method of Bicine, just mix HEPES and NaOH solution in proportion.


The preparation method of EPPS buffer:

(1) With 0.1M solution (A): EPPS25.233g/ deionized water 1,000ml

(2) With 0.1M NaOH solution (B): NaOH4g/ deionized water 1,000ml

(3) Mix the two solutions A and B in proportion and adjust according to the pH value


It can be seen that the three buffers of Bicine, HEPES, and EPPS are all similar in their preparation methods. They all use sodium hydroxide lye to adjust the pH value. Mix in proportion to obtain a buffer solution with a specific pH value. If you need a more accurate pH value, you need to measure the pH and adjust the mixing ratio as needed.


It should be noted that: the preparation temperature is controlled at about 20 degrees; if the detection environment needs to exclude sodium ions, it is also possible to replace sodium hydroxide with potassium hydroxide, and change the weighing mass according to the corresponding molecular weight. Desheng is a manufacturer specializing in the production of buffer raw materials, providing a variety of biological buffer raw materials including Bicine, HEPES and EPPS.