Why is edta added to MOPS buffer?

MOPS (3-morpholinepropanesulfonic acid) is a common buffer that is widely used in many biochemical and molecular biology experiments. However, in some experiments, we may notice the practice of adding EDTA (ethylenediaminetetraacetic acid) to MOPS buffer, what is going on? The following will specifically take you through the reasons why EDTA is added to MOPS buffer, and discuss its role and advantages.

I. MOPS buffer and EDTA

MOPS is an amphoteric buffer, which can maintain a stable pH range of the solution, and it is widely used in biochemical and molecular biology experiments, such as protein electrophoresis and nucleic acid analysis. And EDTA is a chelating agent with strong ability to chelate metal ions, which can form stable complexes with metal ions, thus preventing the interference of metal ions to experiments.

Second, why add EDTA in MOPS buffer

1, metal ion chelation: In some experiments, especially those involving metal ion sensitive experiments, such as enzyme activity assay or nucleic acid analysis, the presence of metal ions may interfere with the experimental results. By adding EDTA, it can form complexes with metal ions, stabilize the metal away from the substance, thus preventing it from affecting the experiment, which can to some extent ensure the accuracy and reproducibility of the experimental results.

2, to prevent metal-catalyzed oxidation reactions: certain metal ions, such as iron ions may be involved in oxidation reactions, and lead to oxidative damage to experimental samples. the role of EDTA can effectively combine with metal ions to inhibit metal-catalyzed oxidation reactions, thereby protecting the integrity of experimental samples.

3, maintain the stability of the buffer: the addition of EDTA can maintain the stability of MOPS buffer. In some experimental conditions, such as long storage or high-temperature treatment, the presence of metal ions may lead to the instability of the buffer, through EDTA can reduce the activity of metal ions, slowing down the degradation and oxidation process of the solution.

Third, the use of EDTA precautions

1, the appropriate concentration: the addition of EDTA should be optimized according to the specific experimental requirements, too high a concentration of EDTA may have a negative impact on some experiments, such as the inhibition of enzyme activity.

2, avoid metal contamination: EDTA should use high-purity preparations, and avoid contact with metal ion contaminated reagents or containers to prevent the introduction of additional metal ions into the experimental system.

3, pH adjustment: the chelating ability of EDTA for different metal ions is affected by pH, so proper pH adjustment needs to be considered in the process of use.

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