| Size | Price | Stock | Qty |
|---|---|---|---|
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg | |||
| Other Sizes |
| ln Vitro |
1. Cytoprotective activity against H₂O₂-induced damage in HeLa cells: Phosphoenolpyruvic acid tricyclohexylammonium salt (providing phosphoenolpyruvic acid, PEP) exerted cytoprotective effects. When HeLa cells were pretreated with the compound at concentrations of 0.1 mM, 0.5 mM, 1 mM, 2 mM, and 5 mM for 24 hours, followed by exposure to 200 μM H₂O₂ for 2 hours, the cell viability (measured by MTT assay) increased in a concentration-dependent manner. At 2 mM, the cell survival rate was significantly elevated to approximately 78%, compared to 42% in the H₂O₂-only control group [1]
2. Antioxidant activity: The compound reduced intracellular reactive oxygen species (ROS) levels in H₂O₂-treated HeLa cells. Pretreatment with 2 mM Phosphoenolpyruvic acid tricyclohexylammonium salt decreased ROS accumulation by about 52% (detected via DCFH-DA fluorescent probe) compared to the H₂O₂-only group. Additionally, it increased the activity of glutathione peroxidase (GSH-Px) by approximately 45% in HeLa cells, enhancing the cellular antioxidant defense system [1] 3. No anti-proliferative activity on normal HeLa cells: Treatment with Phosphoenolpyruvic acid tricyclohexylammonium salt alone (0.1-5 mM) for 24 hours did not affect the viability of non-stressed HeLa cells, with cell survival rates maintained above 95%, indicating no cytotoxicity under normal conditions [1] |
|---|---|
| Enzyme Assay |
1. Glutathione peroxidase (GSH-Px) activity assay: HeLa cells were lysed after treatment with Phosphoenolpyruvic acid tricyclohexylammonium salt (2 mM) and H₂O₂. The cell lysate was mixed with a reaction buffer containing reduced glutathione (GSH), hydrogen peroxide (H₂O₂), and sodium azide. The mixture was incubated at 37°C for 5 minutes, then 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) solution was added to detect the remaining GSH. The absorbance was measured at 412 nm, and GSH-Px activity was calculated based on the rate of GSH consumption, with activity expressed as units per milligram of protein [1]
2. Intracellular ROS detection assay: HeLa cells were loaded with 10 μM DCFH-DA fluorescent probe for 30 minutes at 37°C after pretreatment with Phosphoenolpyruvic acid tricyclohexylammonium salt (0.5-2 mM) and H₂O₂ exposure. The cells were then washed with PBS to remove unloaded probe, and the fluorescence intensity (excitation: 488 nm, emission: 525 nm) was measured using a flow cytometer. ROS levels were quantified by comparing the mean fluorescence intensity of the treated group with the control group [1] |
| Cell Assay |
1. MTT cell viability assay: HeLa cells were seeded in 96-well plates at a density of 5×10³ cells per well and cultured overnight. The cells were then pretreated with Phosphoenolpyruvic acid tricyclohexylammonium salt (0.1-5 mM) for 24 hours, followed by 200 μM H₂O₂ for 2 hours. After treatment, 20 μL of MTT solution (5 mg/mL) was added to each well, and the plates were incubated at 37°C for 4 hours. The supernatant was removed, and 150 μL of dimethyl sulfoxide (DMSO) was added to dissolve the formazan crystals. The absorbance was measured at 570 nm using a microplate reader, and cell viability was calculated as a percentage relative to the untreated control group [1]
2. Hoechst 33342 apoptosis staining assay: HeLa cells were grown on coverslips and treated with Phosphoenolpyruvic acid tricyclohexylammonium salt (2 mM) for 24 hours, then exposed to 200 μM H₂O₂ for 2 hours. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes, and stained with 10 μg/mL Hoechst 33342 for 10 minutes at room temperature. Apoptotic cells (characterized by condensed and fragmented nuclei) were observed under a fluorescence microscope, and the apoptosis rate was calculated by counting apoptotic cells among 300 total cells in five random fields [1] |
| References | |
| Additional Infomation |
Phosphoenolpyruvate tricyclohexylammonium is a salt form of phosphoenolpyruvate (PEP), an intermediate metabolite of glycolysis. This study used this salt form to deliver PEP to HeLa cells because PEP itself has poor cell membrane permeability [1]. The cell protection and antioxidant effects of phosphoenolpyruvate tricyclohexylammonium are achieved by maintaining intracellular redox balance—specifically, by enhancing GSH-Px activity to scavenge excess reactive oxygen species (ROS) and reduce oxidative stress-induced cell damage [1].
|
| Molecular Formula |
C3H2O6P-3.3[C6H14N+]
|
|---|---|
| Molecular Weight |
465.56436
|
| Exact Mass |
465.296
|
| CAS # |
35556-70-8
|
| Related CAS # |
Phosphoenolpyruvic acid potassium;4265-07-0
|
| PubChem CID |
11754241
|
| Appearance |
White to off-white solid powder
|
| Boiling Point |
466.7ºC at 760 mmHg
|
| Melting Point |
197-198 °C
|
| Flash Point |
236.1ºC
|
| LogP |
0.918
|
| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
31
|
| Complexity |
247
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
MJKYGUXBFYGLLM-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/3C6H13N.C3H5O6P/c3*7-6-4-2-1-3-5-6;1-2(3(4)5)9-10(6,7)8/h3*6H,1-5,7H2;1H2,(H,4,5)(H2,6,7,8)
|
| Chemical Name |
cyclohexanamine;2-phosphonooxyprop-2-enoic acid
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
H2O : ~100 mg/mL (~214.80 mM)
DMSO :< 1 mg/mL |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 50 mg/mL (107.40 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1480 mL | 10.7398 mL | 21.4795 mL | |
| 5 mM | 0.4296 mL | 2.1480 mL | 4.2959 mL | |
| 10 mM | 0.2148 mL | 1.0740 mL | 2.1480 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
