Product Name
Phospho(enol)pyruvic acid cyclohexylammonium salt, 98%
Certificate of Analysis (COA)
Product Introduction
Phospho(enol)pyruvic acid cyclohexylammonium salt (PEP salt) is a stabilized salt form of phosphoenolpyruvate (PEP), a key high-energy intermediate in the glycolysis pathway. As an efficient phosphate donor, PEP is widely used in pyruvate kinase assays, ATP regeneration systems, enzyme-coupled kinetic reactions, and metabolic research.
This product is particularly suitable for:
- Pyruvate kinase (PK) activity assays
- PK/LDH coupled enzyme systems
- ATP regeneration in in vitro transcription/translation
- Kinase activity screening
- Metabolic pathway analysis
- Drug discovery and inhibitor screening
The cyclohexylammonium salt form offers improved stability and handling convenience compared to free acid forms.
Mechanism of Action
PEP functions as a high-energy phosphate donor in biochemical systems.
Primary Reaction (Pyruvate Kinase Reaction)
PEP + ADP → Pyruvate + ATP
Coupled Detection System (PK/LDH System)
Pyruvate + NADH → Lactate + NAD⁺
- NADH absorbance decreases at 340 nm
- The rate of NADH consumption is proportional to enzyme activity
- Enables continuous spectrophotometric monitoring
This makes PEP a standard substrate in 340 nm kinetic assays.
Typical Applications
1.Enzyme-Coupled Kinetic Assays
- NADH-based spectrophotometric assays
- Continuous reaction monitoring
- High-throughput microplate assays (96/384 well)
- Drug discovery screening platforms
2.Diagnostic & IVD Reagent Development
- Clinical enzyme detection reagent
- Automated biochemical analyzer systems
- Metabolic disease biomarker studies
3.Pyruvate Kinase (PK) Activity Assays
- Km and Vmax determination
- Enzyme mutation studies
- Metabolic regulation analysis
- PK inhibitor screening
- Cancer metabolism research
4.ATP Regeneration Systems
PEP is widely used as an ATP-regenerating substrate in:
- Cell-free protein synthesis
- In vitro transcription systems
- Kinase cascade assays
- CRISPR and molecular biology reactions
- Signal transduction studies
5.Metabolic & Systems Biology Research
- Glycolytic flux studies
- Energy metabolism modeling
- Systems-level enzyme network analysis
- Biomarker enzyme research
Advantages in Enzyme Assays
- High-energy phosphate donor with well-established kinetics
- Reliable 340 nm spectrophotometric detection
- Suitable for continuous monitoring
- Compatible with PK/LDH coupled systems
- Ideal for ATP regeneration applications
- Stable cyclohexylammonium salt form
- Cost-effective for large-scale screening
- Literature-validated substrate in metabolic research
Recommended Experimental Conditions
| Parameter |
Typical Condition |
| Detection wavelength |
340 nm (NADH-based coupled assay) |
| Detection type |
Continuous kinetic readout |
| Buffer systems |
Tris-HCl, HEPES, Phosphate |
| Required cofactors |
Mg²⁺, K⁺, ADP |
| Optimal pH |
7.0–8.0 |
| Reaction temperature |
25–37°C |
| Assay format |
Spectrophotometric, Microplate, ATP regeneration systems |
Related Research Areas
Researchers working in the following areas may benefit from this product:
- Glycolysis and metabolic pathway research
- ATP regeneration systems
- Kinase activity assays
- Enzyme kinetics studies
- Drug discovery and metabolic inhibitor screening
- Cell-free protein expression systems
