Gluconeogenesis Analysis Service

What is Gluconeogenesis?

Gluconeogenesis is a central metabolic pathway in which glucose is synthesized de novo from non-carbohydrate substrates such as lactate, glycerol, and glucogenic amino acids. This process is crucial for maintaining blood glucose levels during prolonged fasting, intensive exercise, or carbohydrate-restricted conditions. Predominantly occurring in the liver and kidneys, gluconeogenesis is tightly regulated and interconnected with other key metabolic pathways, including glycolysis, the tricarboxylic acid (TCA) cycle, and amino acid metabolism.

Creative Proteomics's gluconeogenesis analysis service is designed to deliver high-resolution, quantifiable, and reproducible insights into this complex pathway. Utilizing state-of-the-art analytical platforms and bioinformatics, we enable researchers to investigate the flux, regulation, and interaction of gluconeogenic intermediates and enzymes under various physiological and experimental conditions.

Advantages of Gluconeogenesis Analysis Service

• Quantitative Accuracy: Our isotope-labeled tracing methods provide quantitation with CVs <10% across biological replicates, enabling precise flux measurements.
• High Throughput Capability: Analyze up to 200 samples per run with batch processing to meet high-volume project needs.
• Wide Dynamic Range: Detection range of 10 nM to 100 µM, ensuring both trace and abundant metabolites are captured effectively.
• Pathway-Integrated Interpretation: Integrated metabolic modeling and pathway visualization support downstream interpretation and hypothesis generation.
• Time-Resolved Kinetics: Kinetic analysis with time-course sampling from 5 min to 24 h post-substrate administration.
• Cross-Species Compatibility: Validated in human, mouse, rat, and other model organisms for translational research.

Gluconeogenesis Analysis Service Offered by Creative Proteomics

• Stable isotope-resolved metabolomics using 13C/15N tracers
• Quantitative flux analysis through the gluconeogenesis pathway
• Targeted quantification of key gluconeogenesis intermediates via LC-MS/MS and GC-MS
• Enzyme activity profiling for PEPCK, FBPase, G6Pase, and related enzymes
• Time-course sampling and dynamic metabolite profiling
• Comprehensive mapping of gluconeogenesis and related metabolic pathways
• Visualization of metabolic shifts through custom pathway diagrams
• Customized statistical analysis and biological interpretation
• Multi-omics data integration (metabolomics + transcriptomics/proteomics)
• Species-specific method customization and tracer adaptation

List of Detected Gluconeogenesis and Related Metabolites

Core Gluconeogenesis Intermediates

Connected TCA Cycle Intermediates

Amino Acid–Derived Gluconeogenic Substrates

Lipid and Glycerol Pathway Intermediates

Energy and Redox State Indicators

Workflow for Gluconeogenesis Analysis Service

Technology Platform for Gluconeogenesis Analysis Service

Sample Requirements for Gluconeogenesis Analysis Service

Applications of Gluconeogenesis Assay Service

LC-MS/MS analysis of the PGB 1 glucuronide biosynthesized by HL microsomes (Little, Joanna M., et al., 2004).

Reference

1. Little, Joanna M., et al. "Glucuronidation of oxidized fatty acids and prostaglandins B1 and E2 by human hepatic and recombinant UDP-glucuronosyltransferases." Journal of lipid research 45.9 (2004): 1694-1703. https://doi.org/10.1194/jlr.M400103-JLR200

Q1: Can I analyze both intracellular and extracellular metabolites in one experiment?

A1: Yes. We offer separate or combined profiling of intracellular and extracellular fractions, depending on your study design. Please specify during consultation.

Q2: Do you support isotope-labeled tracer optimization for custom substrates?

A2: Yes. We provide consultation on tracer selection and labeling strategies, including custom tracers like [U-13C] glycerol or [2-13C] pyruvate.

Q3: What normalization methods are used for metabolite quantification?

A3: We offer normalization by cell number, protein content, tissue weight, or external/internal standards, based on your study needs.

Q4: Can I submit samples in organic solvents (e.g., methanol extract)?

A4: Yes. We accept both raw biological samples and pre-extracted metabolites. Please contact us for solvent compatibility and volume requirements.

Q5: What if I only have a limited sample amount?

A5: We can work with minimal sample input and offer pre-analysis consultation to adjust detection strategies for low-volume samples.

Q6: Do you provide flux calculation using isotope labeling data?

A6: Yes. We offer metabolic flux analysis (MFA) including isotopomer distribution models and pathway-specific flux quantification.

Q7: Can you help design my gluconeogenesis experiment from scratch?

A7: Absolutely. We provide end-to-end experimental design support, including controls, sampling time points, and data interpretation strategy.

Q8: Is batch effect correction included in your data analysis?

A8: Yes. Our pipeline includes statistical correction for batch effects, ensuring high comparability across sample groups.

Q9: Will I receive pathway visualization in the report?

A9: Yes. Our standard report includes annotated pathway maps, flux diagrams (if applicable), and detailed metabolite heatmaps.

Q10: Can you perform analysis on archived or long-stored samples?

A10: Yes, provided that the samples have been stored properly at -80°C without multiple freeze-thaw cycles. We can help evaluate sample integrity.

Q11: What is the detection limit of the Agilent 6495C Triple Quadrupole for gluconeogenic intermediates?

A11: The Agilent 6495C LC/MS system achieves a detection limit of 0.1 fmol/μL for key metabolites like phosphoenolpyruvate (PEP) and oxaloacetate, with a linear dynamic range spanning six orders of magnitude (R² > 0.998).

Q12: How does the Agilent 7890A GC ensure reproducibility for glycerol quantification?

A12: Using the VF-WAXms column (30 m × 0.25 mm), we achieve a retention time CV <0.5% for glycerol in plasma samples. The system's electron impact ionization (EI) mode ensures 99% isotopic purity for [2H]-labeled tracers