There are many cases where it is not practical or possible to label the experimental sample, for example, when working with tissue samples, performing large fermentation runs, or propagating field-grown plants. In these instances, the IROA Phenotypic protocol may be applied. In the Phenotypic IROA protocol (Figure 3-3), the control or “Internal Standard” is isotopically labeled using U-13C 95% IROA media.
An ideal Internal Standard is one that represents the entire metabolome of the fluid or tissue under study. For example, if interrogating the metabolome in liver tissue, quantification of metabolites can be achieved bu growing and labeling a liver cell line to use as a complex “Internal Standard”. For Phenotypic analyses, ClusterFinder software will find all IROA-labeled compounds/peaks in the Internal Standard and use these 13C IROA peaks to identify their associated natural abundance peaks in the experimental sample.
The IROA Phenotypic Quantitation Kit for Mammalian Metabolic Profiling is designed to create a labeled complex Control or Internal Standard to spike into experimental samples.
IROA PHENO-95-300 KIT: $2,240.00
Materials: EBSS/RPMI 1640 vitamins PLUS U-13C, 95% labeled carbon energy sources; ClusterFinder software