Spectrophotometric analysis, also called spectral scanning, of biomolecules serves two main purposes – the quantitation of nucleic acids (DNA and RNA) and purity assessment. Since the amount or concentration and purity of the DNA or RNA in a solution significantly affect the results of subsequent downstream applications, establishing these values early on can reduce, if not totally eliminate, the risk of committing errors and guarantee optimum results.
Various types of commercial-grade detergents contain elevated levels of sulfhydryl oxidizing agents, peroxides, salts, and carbonyl compounds. The proteins that are isolated by these detergents are highly susceptible to contaminating peroxides and carbonyls. The peroxides oxidize the proteins, and the carbonyl groups will form Schiff’s bases with the proteins that interfere with its structure.
As a non-ionic viscous liquid, tweens are a class of emulsifiers used in pharmaceuticals and food preparation.
While detergents play a vital role in separating proteins from the hydrophobic portions of the cell membrane during extraction and sample preparation, they need to be removed after successfully serving their purpose to prevent them from interfering with downstream applications (e.g., IEF, ELISA, protease digestion of proteins).