Buffer solution is used in biochemical experiments to stabilize the pH of the system. MOPS, 3-(N-Morpholino)Propanesulfonic Acid, MES, 2-Morpholinoethanesulfonic acid, are commonly used in biological experiments, and played very important role. What is the difference between the two buffers? How to prepare the buffer solution?
MOPS (CAS 1132-61-2), 3-(N-Morpholino)Propanesulfonic Acid
The buffer range of MOPS is 6.5~7.9. It can be used:
(1) As running buffer in electrophoresis and for protein purification in chromatography;
(2) As Lysis buffer for Escherichia coli cells;
(3) As buffer for RNA isolation and transfection in Northern blot;
(4) In agar medium for the culture of bacteria, yeast and mammalian cells;
(5) As eluent in gel filtration chromatography;
(6) For bicinchoninic acid (BCA) assay;
(7) For study on electron transport and phosphorylation of chloroplast sample preparation.
Preparation method for 10 × MOPS Buffer is as follows:
(1) 41.8g MOPS is dissolved in about 700mL DEPC treated water;
(2) Adjusted pH to 7.0 with 2N NaOH;
(3) 20mL of 1M NaOAC and 0.5M EDTA (pH 8.0) treated with DEPC were added to the solution;
(4) Make the volume to 1L, then filter with 0.45μm filter membrane to remove impurities, store at room temperature and protect from light.
MES Buffer, 2-Morpholineethanesulfonic Acid
The buffer range is 5.5~6.7, pKa value of MES buffer is near to physiological pH, it is used in the following experiments:
(1) Replace highly toxic cacodylate, and ion buffer citrate and malate;
(2) Buffered media for bacterial, yeast and mammalian cells. High concentrations of MES are toxic to most plants, but can be used in plant media at concentrations ~10 mM;
(3) Studying Tau protein;
(4) Electrophoresis and chromatography including capillary electrochromatography, gel filtration chromatography, phosphocellulose column chromatography, hydrophobic interaction chromatography, cation exchange chromatography and SDS-PAGE;
(5) Complex with organotin (IV) molecules as antitumor agents.
Preparation method is as follows:
Assume that the pH is 6 and the concentration is 100mM:
(1) 0.1mol or 19.5g of MES was weighed and dissolved in 500mL of DEPC water;
(2) The pH was adjusted to 6.0 with 1N or 10N NaOH solution;
(3) Make the volume to 1L, preserve at 4°C.
The choice of buffer is not only according to the required buffer range and capacity of buffer solution, but also according to the specific experimental content. In addition, the experimenter should pay attention to the sterilization of containers, water and other additives.
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