Description
Abbreviated as Tris HCl (1185-53-1), Tris (hydroxymethyl) aminomethane hydrochloride is an organic compound commonly utilized in buffer solutions such as TAE or TBE for electrophoresis gels. It boasts high solubility in water and offers utility within the pH range of 7.0-9.0. The compound is instrumental in preparing Laemmli buffer, one of the most widely used SDS-PAGE buffers, and also finds application in various custom running and loading buffers, frequently in combination with glycine and SDS. Lysis, the initial step of DNA extraction, involves a buffer containing Tris and EDTA (ethylenediaminetetraacetic acid). In this process, EDTA binds to divalent cations such as calcium and magnesium. Due to their role in maintaining cell membrane integrity, their removal by EDTA leads to membrane destabilization. Tris plays a key role as the primary buffering component, maintaining the pH of the buffer at a stable point, typically around 8.0. Additionally, Tris may interact with the LPS (lipopolysaccharide) in the membrane, contributing to further membrane destabilization. This underscores Tris HCl's significance in various laboratory procedures.
Chemical Properties
White crystalline powder
Uses
For precise applications, use a carefully calibrated pH meter with a glass/calomel combination electrode;For Tris buffers, pH increases about 0.03 unit/°C decrease in temperature, and decreases 0.03-0.05 unit per ten-fold dilution.
Application
TRIS hydrochloride is highly soluble in water and is useful in the pH range 7.0-9.0. It is used in the preparation of Laemmli buffer, one of the most common SDS-PAGE buffers. In addition, Tris can also be used for many custom running and loading buffers, most often with glycine and SDS. The use of single-junction pH electrodes that contain silver should be avoided when determining the pH of a solution containing Tris buffer.
Preparation
The preparation of TRIS hydrochloride is as follows:After reacting 2-amino-1,3-propanediol (2.50 g, 27.44 mmol) and formic acid (1.26 g, 27.44 mmol) in 50 ml of water at room temperature for 3 hours, water was distilled off under reduced pressure, A liquid was obtained. By washing the obtained liquid,An ammonium formate salt (3.76 g, 27.44 mmol) was obtained as a colorless transparent liquid.
General Description
Trizma? hydrochloride is mainly required for preparation of buffer at physiological range of 7.3 to 7.5. The prepared buffer is compatible with biological fluids. It is of importance to laboratories as a standard pH solution.
Biological Activity
Commonly used laboratory reagent
Purification Methods
Crystallise the salt from 50% EtOH, then from 70% EtOH. TRIS-hydrochloride is also available commercially in a highly pure state. Otherwise, recrystallise it from 50% EtOH, then 70% EtOH, and dry it below 40o to avoid risk of decomposition. [Beilstein 4 H 304.]
