Quantitative determination of fluorescence labeling implemented in cell cultures.

ABSTRACT:

Background

Labeling efficiency is a crucial parameter in fluorescence applications, especially when studying biomolecular interactions. Current approaches for estimating the yield of fluorescent labeling have critical drawbacks that usually lead them to be inaccurate or not quantitative.

Results

We present a method to quantify fluorescent-labeling efficiency that addresses the critical issues marring existing approaches. The method operates in the same conditions of the target experiments by exploiting a ratiometric evaluation with two fluorophores used in sequential reactions. We show the ability of the protocol to extract reliable quantification for different fluorescent probes, reagents concentrations, and reaction timing and to optimize labeling performance. As paradigm,

SUBMITTER: Schirripa Spagnolo C 

PROVIDER: S-EPMC10496409 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature