Biological buffer is a kind of solution that can keep the pH value of the solution relatively stable when a small amount of acid or alkali is added. Most cells can only carry out activities in a very narrow pH range, and need a buffer system to resist the pH changes in the process of metabolism. In biochemical research, buffer solution is often used to maintain the pH of the experimental system. Let's take a look at the commonly used biological buffer solutions and their characteristics.
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Phosphate buffer
Phosphate buffer is the most widely used buffer. Due to the secondary dissociation, the pH range of the buffer is wide, and the acidic, alkaline and neutral buffers with different pH values can be configured
When preparing acidic buffer solution, NaH2PO4 or KH2PO4 can be used directly, and the pH value ranges from 1 to 5;
Alkaline buffer can be directly used Na2HPO4 or K2HPO4 with pH range of 9-12;
The neutral buffer contains the same amount of NaH2PO4 and Na2HPO4 or the same amount of KH2PO4 and K2HPO4 solution, and the pH value is 5.5 ~ 8.5.
Advantages: ① easy to prepare into various concentrations; ② wide range of pH value; ③ little influence of temperature on pH value;
Disadvantages: ① it is easy to precipitate with common calcium, magnesium and heavy metal ions; ② it inhibits some biochemical processes;
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Tris buffer
Tris buffer is widely used in biochemical research. It is a weak base and is usually used in the "neutral" range. In addition to tris - HCl, Tris has a variety of derivatization buffers
TBS = Tris HCl + NaCl + KCl, which is often used to clean the Western blotting membrane of immunostained tissue or Western blotting;
Tbst = Tris HCl + NaCl + Tween20, a membrane buffer commonly used for Western blotting;
TE = Tris HCl + EDTA, which has protective effect on DNA base and is often used for DNA stabilization and storage;
Tae = Tris base + acetic acid + EDTA is a buffer system widely used in short DNA electrophoresis;
Tbe = tribasic + boric acid + EDTA, suitable for long-term DNA electrophoresis, with good separation effect for small fragments
Advantages: ① because Tris base has strong alkalinity, it can only be used to prepare buffer solution with a wide range of pH from acidic to alkaline; ② it has little interference to biochemical process and does not precipitate with calcium, magnesium ions and heavy metal ions.
Disadvantages: ① the pH value of the buffer solution is greatly affected by the concentration of the solution, and the pH value changes more than 0.1 when the buffer solution is diluted ten times; ② the temperature effect is large, and the temperature change has a great influence on the pH value of the buffer solution, 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. ④ The buffer solution interferes with some pH electrodes, so the electrode compatible with tris solution should be used.
Glycine buffer
Glycine buffer has a wide range of pH value and wide range of application, ranging from 2.0 to 11.0.
Advantages: provide a closer natural environment for cell components and various extracts.
Disadvantages: similar to phosphate buffer system; interferes with some biochemical processes, such as metabolism.
Desheng technology was founded in 2005. After years of innovation and development, the company has become a national high-tech enterprise with R & D, production, sales and service as a whole, which is dominated by in vitro diagnostic reagent raw materials, biological buffer, blood collection additive, color reagent and chemiluminescence reagent.