The role of trihydroxymethyl aminomethane 77-86-1 in Western blot experiments

Western blot is a classic technique in molecular biology for detecting specific proteins, which involves multiple steps such as sample processing, electrophoretic separation, transfer printing, and antibody detection. Trihydroxymethyl aminomethane, as a core buffering agent, provides critical support for the accuracy and reproducibility of experiments by maintaining a stable pH environment, optimizing ion strength, and protecting protein activity.

1, Stable pH environment, protecting protein structure

The charge distribution, solubility, and antigen epitope exposure of proteins are highly dependent on pH value. The buffering range of trihydroxymethylaminomethane (pH 7.0-9.0) is highly compatible with the commonly used conditions in Western blot experiments, and can effectively resist pH fluctuations caused by the addition of chemical reagents or electrochemical reactions during the experimental process.

trihydroxymethylaminomethane maintains powder

During the sample lysis stage, the lysis buffer containing trihydroxymethylaminomethane maintains a neutral to weakly alkaline environment, inhibits endogenous protease activity, and prevents degradation of the target protein. In the preparation of SDS-PAGE gel, trimethylolaminomethane, acrylamide, ammonium persulfate and other components jointly build a stable polymerization system, whose pH stability ensures that the pore diameter of the gel is uniform, and avoids precipitation or conformational change of protein due to local acidification. In addition, the trihydroxymethylaminomethane in the transfer buffer neutralizes the surface charge of the membrane, reduces non-specific binding between the protein and the membrane, and further protects the protein antigenicity.

2, Optimize the electrophoresis system to achieve precise separation

The concentration design of trihydroxymethylaminomethane has been optimized: too low a concentration can lead to insufficient conductivity and unstable current; If the concentration is too high, it may cause gel heating or band diffusion. Its moderate ionic strength can not only ensure the electrophoresis efficiency, but also maintain the stability of gel pore size, so that different molecular weight proteins can be separated clearly.

3, Promote transfer efficiency and ensure complete protein migration

Maintaining ion balance: Tris (hydroxymethyl) aminomethane, glycine, and methanol together form a transfer printing solution. Its pH stability (usually pH 8.3) ensures that the protein maintains its natural conformation during the transfer printing process, avoiding denaturation or precipitation.

Optimizing electric field driving: The trihydroxymethyl aminomethane glycine system forms a stable electric field gradient, driving proteins to migrate towards the membrane at a constant rate. Methanol can destroy the gel network structure, while trimethylolaminomethane can balance the ionic strength through buffering, prevent the excessive accumulation of membrane surface charges, and thus improve the transfer efficiency of large molecular weight proteins.

Protective membrane protein: The mild chemical properties of trihydroxymethylaminomethane prevent the membrane protein from being damaged by strong acids or bases, ensuring the complete exposure of antigen epitopes, thereby enhancing the sensitivity and accuracy of antibody binding.

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Hubei Xindesheng Material Technology Co., Ltd. has been specializing in the research and production of biological buffering agents for nearly 20 years. We have unique insights into the selection of raw materials, updates in production technology, product quality testing, and control of production costs. We are committed to providing customers with affordable and high-quality products! At present, Desheng Company can provide dozens of buffering agents such as trihydroxymethylaminomethane. If you have any related purchasing needs in the near future, please click on the official website to learn more details or contact me directly!