The IROA 300 Mammalian Metabolic Profiling Kit is a simple and direct metabolic labeling protocol for the in-vivo incorporation of specific and unique signatures or tags into biochemical metabolites for relative quantitation by mass spectrometry (MS) and can be applied to many life science applications (toxicology, diagnostics, drug development, bioprocess, etc.).
The IROA 300-250 Kit is designed to label Control cell populations with 95% U-13C media and Experimental cell populations with 5% U-13C media, “basic” profiling.
IROA 300-250 is supplied as (2) Kits:
1) IROA PHENO-95-300 Kit (labeling kit for Control cell populations) and, 2) the IROA FLUX-05-300 Kit (labeling kit for Experimental cell populations).
Note: These kits can also be used independently for “phenotypic” analysis (PHENO-95-300: when comparing unlabeled natural abundance Experimental cells vs. labeled Control cells) or for “fluxomic” profiling. The PHENO and FLUXOMIC kits are combined (IROA 300-250 for “basic” profiling.
IROA Mammalian Metabolic Profiling Kit- Cat No. IROA 300-250: $3594.00
Materials: EBSS/RPMI 1640 vitamins PLUS U-13C, 95% and 5% labeled carbon energy sources; ClusterFinder software
IROA PHENO-95-300 KIT (ONLY): $2,240.00
Materials: EBSS/RPMI 1640 vitamins PLUS U-13C, 95% labeled carbon energy sources; ClusterFinder software
Additional Materials Required
- Unlabeled IROA Mammalian medium, IROA Cat. No. IROA-300-UL; for the adaption of cells to IROA media
- Dialyzed Fetal Bovine Serum (dFBS); IROA Cat. No. 300-DS-75
- VWR Vacuum filtration system, PES 0.2 µm, 250mL bottle, sterile; VWR Cat # 97066-200 or other suitable filtration system
- Phosphate-buffered saline (PBS): 0.01 M; pH 7.4; Sigma Cat. No. P5368-10PAK, or equivalent
- Bradford reagent, Sigma Cat. No, B6916-500ML, for protein determination, optional
To ensure that the IROA media supports the growth and proliferation of your cells, it is recommended that cells are initially grown and tested in IROA unlabeled mammalian media, (IROA Cat. No. IROA-300-UL, $200.00), prior to labeling with the Mammalian profiling kit. At least 5 or more cell doubling cycles are required to achieve full label incorporation.