2-nitrophenyl β-D-galactopyranoside-[1-13C]

2-Nitrophenyl β-D-galactopyranoside-[1-13C], often referred to as ONPG-[1-13C], serves as a fundamental substrate in biochemical and molecular biology research. Here are the key applications of this compound:

Enzyme Activity Assays: Central to the realm of biochemical investigations, ONPG-[1-13C] acts as a widely adopted chromogenic substrate for assessing the activity of β-galactosidase, an enzyme tasked with hydrolyzing β-galactosides into monosaccharides. Upon interaction with β-galactosidase, ONPG-[1-13C] unleashes a vivid yellow product – 2-nitrophenol – allowing for precise quantitative measurements. This analytical technique empowers researchers to unravel enzyme kinetics and delve into the intricate mechanisms governing enzymatic actions.

Isotopic Tracing: Leveraging the distinctive 13C isotope label embedded within ONPG-[1-13C], this compound emerges as a pivotal tool for conducting isotopic tracing studies within the realm of metabolic exploration. By meticulously monitoring the assimilation and conversion of the 13C label, scientists can delve into the metabolic pathways involving β-galactosides, shedding light on the nuances of carbohydrate metabolism and its elaborate regulatory networks across diverse biological contexts.

Reporter Gene Assays: A cornerstone in gene expression analyses, ONPG-[1-13C] finds widespread utility in reporter gene assays aimed at exploring gene expression patterns and promoter activities. Particularly when the lacZ gene, encoding β-galactosidase, is utilized as a reporter, the enzymatic breakdown of ONPG-[1-13C] serves as a quantitative indicator of gene expression levels. This methodological approach proves invaluable for unraveling the intricacies of gene regulation and discerning the impacts of genetic alterations on cellular functions.

Microbial Identification: In the field of microbiology, the deployment of ONPG-[1-13C] stands pivotal for discriminating between lactose-fermenting and non-fermenting bacterial strains. The capacity of microorganisms to metabolize ONPG-[1-13C], indicative of the presence of β-galactosidase, serves as a diagnostic beacon for microbial identification purposes. This diagnostic modality proves particularly instrumental in clinical settings, allowing for rapid and accurate identification of pathogenic bacterial species, thereby facilitating targeted therapeutic interventions.