Half-life Assay by Pulse-chase

The amount of time it takes for a drug’s concentration in the body to drop to 50% is known as its half-life. The drug’s half-life provides insight into how the drug is metabolized within the body. It is a crucial pharmacokinetics parameter that also serves as a reliable measure of the rate at which drugs are eliminated from the body, which is what is used to establish the dosage and frequency of administration.

An established and incredibly flexible method for researching the life cycle of drugs is pulse-chase analysis. Given that it has no effect on normal cell development and metabolism, it is also the most widely used technique for determining the half-life of medications, including proteins and peptides. Pulse-chase experiments are divided into two stages: Drugs are exposed to a labeling chemical in the pulse phase, which is integrated into newly synthesized macromolecules of interest;2) The same compound is introduced in excess in the unlabeled form during the second phase, known as the chase phase, to replace the labeled form. No macromolecules will be tagged that are produced during the pursuit phase. Degradation processes cause the quantity of macromolecules generated during the pulse to decrease during the course of the chase. 

We have developed an advanced half-life extension platform based on years of experience. A range of half-life assay is offered in our facility, including pulse-chase assay. Usually, a radioactive precursor is used to metabolically label the target drug for a brief duration. To stop the radiolabel from being further incorporated into medications, an excess of nonradioactive precursor molecules is supplied to the culture during the next chase period. Targeted substances adsorber and precipitate samples at distinct points in the chase period. Lastly, fluorescence analysis is used to quantify the substance that has been radiolabeled.

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