Tris(2 - carboxyethyl)phosphine Hydrochloride, with the CAS number 51805 - 45 - 9, is a well - known reducing agent in the field of biochemistry and molecular biology. It is commonly supplied in a factory - made environment at a low price, which makes it an attractive option for both research institutions and industrial applications.
Chemical Formula: C₉H₁₅O₆P·HCl
Molecular Weight: Approximately 286.65 g/mol
Appearance: Usually a white to off - white crystalline powder
Solubility: Highly soluble in water, which allows for easy preparation of solutions.
Stability: It is relatively stable under normal storage conditions. However, it should be stored in a cool, dry place away from strong oxidizing agents to maintain its quality.
In biochemistry, Tris(2 - carboxyethyl)phosphine Hydrochloride is widely used as a reducing agent. It can effectively break disulfide bonds in proteins and peptides, which is crucial for the analysis and modification of these biomolecules. For example, in protein structure determination experiments, such as X - ray crystallography and NMR spectroscopy, reducing disulfide bonds can help the protein to adopt a more stable and suitable conformation for analysis.
In molecular biology, it is used in DNA and RNA extraction procedures. It can prevent the oxidation of biomolecules during the extraction process, thus maintaining the integrity of the genetic material. Additionally, it is also used in the preparation of antibodies, where reducing disulfide bonds can be necessary for the proper folding and function of the antibody molecules.
In the pharmaceutical industry, it can be used as a stabilizer in the formulation of protein - based drugs. By reducing disulfide bonds, it can prevent protein aggregation and degradation, thereby extending the shelf - life of the drugs.
1. Solution Preparation: Dissolve the appropriate amount of Tris(2 - carboxyethyl)phosphine Hydrochloride in water to prepare a stock solution. The concentration of the stock solution can be adjusted according to the specific application. For most biochemical experiments, a 10 mM - 100 mM stock solution is commonly used.
2. Reaction Conditions: When using it as a reducing agent, the reaction is usually carried out at a pH range of 4 - 8. The reaction temperature can vary from room temperature to 37°C, depending on the specific requirements of the experiment or process.
3. Dosage: The dosage of Tris(2 - carboxyethyl)phosphine Hydrochloride should be determined based on the amount of the target molecule and the reaction conditions. Generally, a molar excess of the reducing agent is used to ensure complete reduction of disulfide bonds.
Case 1: Protein Structure Analysis
A research group was trying to determine the crystal structure of a novel protein. They used Tris(2 - carboxyethyl)phosphine Hydrochloride to reduce the disulfide bonds in the protein. First, they prepared a 50 mM stock solution of the reducing agent. Then, they added an appropriate amount of the stock solution to the protein sample to achieve a final concentration of 10 mM. After incubating the sample at room temperature for 30 minutes, they found that the protein adopted a more stable conformation, which allowed them to obtain high - quality crystals for X - ray crystallography. The crystal structure obtained provided valuable insights into the function of the protein.
Case 2: DNA Extraction
A biotechnology company was performing DNA extraction from plant tissues. They added Tris(2 - carboxyethyl)phosphine Hydrochloride to the extraction buffer at a concentration of 5 mM. By doing so, they were able to prevent the oxidation of DNA during the extraction process. As a result, the extracted DNA had a high purity and integrity, which was suitable for subsequent PCR and sequencing experiments.
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