13C-labeled glutamine for 13C-MFA

13C-labeled glutamine for 13C-MFA

Mutant genes and malfunctioned signaling pathway in cancers affect the regulation of glutamine metabolism. In recent years, glutamine has emerged as a key substrate to support cancer cell proliferation, and the quantification of its metabolic flux is critical to understand the mechanisms by which this amino acid is involved in the metabolism that sustains the survival and growth of tumor cells. Glutamine is not only a major mechanism for nitrogen transport into cells, but also supplements glucose as a substantial carbon source through anaplerosis into the tricarboxylic acid (TCA) cycle. Stable isotope labeling provides a direct readout of intracellular metabolism and can be combined with the known stoichiometry of biochemical pathways to estimate the activity of corresponding enzyme fluxes. Creative Proteomics has employed different labeled glutamine tracers such as [1-¹³C] and [U-¹³C] glutamine to carry out the ¹³C-metabolic flux analysis by generating a quantitative map of cellular metabolism.

Figure 1. [U-13 C] Glutamine tracer experiments produce rich labeling patterns. (Antoniewicz, M. R. 2018)

Our Services

Creative Proteomics has developed a novel metabolic flux analysis platform to provide ¹³C-labeled glutamine for ¹³C-MFA service in a competitive fashion. We can offer a wide range of services to support all research and development activities. Here is our ¹³C-labeled glutamine based metabolic flux analysis procedure

• Design of experiment

¹³C-labeled glutamine tracers are used to trace the catabolism of glutamine carbons in central carbon metabolism. We can perform analysis of extracellular metabolites.

• Cell culture using stable isotopic tracers

The use of [U-¹³C6] glutamine tracer allows metabolites in the TCA cycle to reach isotope stability in a short time and glycolytic metabolites to reach isotope stability in 1.5 hours

• Measurement of extracellular glutamine

Glutamine depletion rate is calculated as a parameter for metabolic flux analysis, a method and metabolic model for estimating intracellular metabolic fluxes based on stable isotopes. Parallel labeling experiments with ¹³C-labeled glutamine substrates are performed to enhance the resolution of metabolic fluxes in different models

• Fluxes quantification in lower metabolism

We have used fully labeled [U-¹³C] glutamine to analyze the lower part of central carbon metabolism, i.e., downstream of pyruvate, specifically in the intermediate products of the TCA cycle

Figure 2. Panel A shows the atom-resolved fate of 50% [1,2-¹³C]-glutamine through glycolysis and the pentose phosphate pathway. (Antoniewicz, M. R. 2018)

Features of Our MFA Platform

• Developed based on the most updated knowledge of biology, bioinformatics and software development
• Widely applicable to a wide range of metabolic system
• Professional bioinformatics teams & personalized bioinformatics analysis services.
• Advanced instrument platform
• Integrated quantitative methodologies and comprehensive solutions for metabolomics

Based on high-performance quantitative techniques and advanced equipment, Creative Proteomics has constantly updating our metabolic flux analysis platform and is committed to offering professional, rapid and high-quality services of ¹³C-labeled glutamine for ¹³C-MFA at competitive prices for global customers. Our personalized and comprehensive services can satisfy any innovative scientific study demands, please contact our specialists to discuss your specific needs. We are looking forward to cooperating with you!

Reference

1. Antoniewicz, M. R. A guide to ¹³C metabolic flux analysis for the cancer biologist. Experimental & Molecular Medicine. 2018. 50: 19.