Dianfeng Chemical
SUPPLEMENTS
2 Mercaptoethanol Western Blot
Application of mercaptoethanol in Western blotting
Blotting is an important method for protein digestion. Among them, 2-mercaptoethanol (2-Mercaptoethanol) is used in Western Blot, which is quite important.
2-mercaptoethanol has strong reducing properties and can break the disulfide bonds within and between molecules of proteins. The protein structure is maintained by the disulfide bond. If it is broken, the protein subunits will dissociate and form a single chain. When processing the sample, add an appropriate amount of 2-mercaptoethanol and incubate it with the protein sample. It quickly enters the protein, breaks the disulfide bond, and makes the protein structure allosteric.
This alteration has various benefits in Western Blot. First, after the dissociation of protein subunits, it is better to separate them according to molecular weight in gel electrophoresis. The swimming rate of each subunit is different, resulting in clear bands, which is convenient for the formation of the molecular weight and subunits of the deprotein. Second, the antigen epitopes are more fully exposed. After the disulfide bond is broken, the hidden epitopes inside the protein appear on the outside, and the antibody binds to it more easily, increasing the sensitivity and specificity of detection.
It is also necessary to pay attention to the use of 2-mercaptoethanol. It is volatile and irritating, and it must be kept in a fume hood during operation, and it must be well protected to avoid harming the body. The dosage should also be accurate. If it is too small, the disulfide bond will be incomplete, which will affect the separation and detection; if it is too much or causes excessive protein denaturation, it will lose its immune activity.
In short, 2-mercaptoethanol plays a key role in Western Blot, and its good use can make the blot results accurate and reliable, making it a good assistant for protein research.
Blotting is an important method for protein digestion. Among them, 2-mercaptoethanol (2-Mercaptoethanol) is used in Western Blot, which is quite important.
2-mercaptoethanol has strong reducing properties and can break the disulfide bonds within and between molecules of proteins. The protein structure is maintained by the disulfide bond. If it is broken, the protein subunits will dissociate and form a single chain. When processing the sample, add an appropriate amount of 2-mercaptoethanol and incubate it with the protein sample. It quickly enters the protein, breaks the disulfide bond, and makes the protein structure allosteric.
This alteration has various benefits in Western Blot. First, after the dissociation of protein subunits, it is better to separate them according to molecular weight in gel electrophoresis. The swimming rate of each subunit is different, resulting in clear bands, which is convenient for the formation of the molecular weight and subunits of the deprotein. Second, the antigen epitopes are more fully exposed. After the disulfide bond is broken, the hidden epitopes inside the protein appear on the outside, and the antibody binds to it more easily, increasing the sensitivity and specificity of detection.
It is also necessary to pay attention to the use of 2-mercaptoethanol. It is volatile and irritating, and it must be kept in a fume hood during operation, and it must be well protected to avoid harming the body. The dosage should also be accurate. If it is too small, the disulfide bond will be incomplete, which will affect the separation and detection; if it is too much or causes excessive protein denaturation, it will lose its immune activity.
In short, 2-mercaptoethanol plays a key role in Western Blot, and its good use can make the blot results accurate and reliable, making it a good assistant for protein research.
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