| Size | Price | |
|---|---|---|
| Other Sizes |
| Targets |
Aminoimidazole carboxamide ribonucleotide transformylase/inosine monophosphate cyclohydrolase (ATIC)[1]
The primary biological targets of DCIP hydrate sodium are enzymes that catalyze reduction-oxidation reactions, including NADH dehydrogenase (Complex I of the mitochondrial electron transport chain), succinate dehydrogenase (Complex II), and various other flavin-dependent oxidoreductases. In photosynthesis research, DCIP acts as an artificial electron acceptor for photosystem II (PSII), accepting electrons from the primary electron acceptor QA. The compound also targets ascorbate (vitamin C) via direct chemical reduction. It does not bind to G protein-coupled receptors or protein kinases; its target class is redox enzymes. |
|---|---|
| ln Vitro |
In vitro activity assays using DCIP measure the rate of reduction of the blue dye to its colorless form, which is monitored spectrophotometrically at 600 nm (ε = 21 mM-¹ cm-¹ at pH 7.0). A decrease in absorbance at 600 nm over time corresponds to reduction. For Complex I (NADH:ubiquinone oxidoreductase), DCIP acts as an artificial electron acceptor: NADH + H+ + DCIP (blue) → NAD+ + DCIPH2 (colorless). Activity is measured in mitochondrial preparations or purified enzyme. For ascorbic acid quantification, the disappearance of the blue color upon mixing with sample is proportional to vitamin C concentration.
|
| ln Vivo |
DCIP hydrate sodium is not used as a therapeutic agent; therefore, in vivo animal studies are not applicable. However, the compound is used in ex vivo tissue assays. For example, in a rat model of ischemia-reperfusion injury, mitochondrial Complex I and II activities can be assessed in heart or brain homogenates using DCIP as an electron acceptor. Isolated mitochondria (50-100 microg protein) are incubated with NADH and DCIP, and the decrease in A600 is measured. Reduced activity correlates with oxidative damage. No pharmacodynamic effects are measured because DCIP is not a drug.
|
| Enzyme Assay |
Standard protocol for measuring NADH dehydrogenase (Complex I) activity using DCIP: Prepare reaction buffer containing 50 mM potassium phosphate (pH 7.5), 1 mM KCN (to inhibit cytochrome c oxidase), 2 microg/mL antimycin A (to inhibit Complex III), and 100 microM DCIP. Add 10-50 microg of mitochondrial protein or 5-25 microg of purified enzyme. Initiate reaction by adding 100 microM NADH (final concentration). Record decrease in absorbance at 600 nm for 2-5 minutes at 25degC. One unit of activity is defined as the reduction of 1 micromol DCIP per minute at 25degC. Include blank without NADH and control with rotenone (10 microM, Complex I inhibitor).
|
| Cell Assay |
DCIP is not typically used in cell viability or proliferation assays because it is a redox dye that interacts with the electron transport chain. However, cell-based assays can measure NADH-driven DCIP reduction. A typical protocol: Plate cells (e.g., HEK293, HepG2) in 96-well plates (20,000 cells/well). The next day, remove media, wash with PBS, add 100 microL of assay buffer (50 mM Tris-HCl pH 7.5, 0.1% Triton X-100, 100 microM DCIP, 1 mM KCN). Add NADH to 100 microM and measure absorbance at 600 nm over 10 minutes using a plate reader. Alternatively, cells can be pre-treated with test compounds (e.g., mitochondrial toxins) to assess effects on dehydrogenase activity. For viability assays, MTT or resazurin (Alamar Blue) is preferred over DCIP.
|
| Animal Protocol |
For animal tissue homogenate analysis: Rats or mice are euthanized, and tissues (e.g., brain, heart, liver, kidney) are rapidly excised and placed in ice-cold isolation buffer (250 mM sucrose, 10 mM Tris-HCl pH 7.4, 1 mM EDTA). Tissues are homogenized, and mitochondria are isolated by differential centrifugation. Mitochondrial pellets are resuspended in buffer and protein concentration determined. For activity measurement, 10-20 microL of mitochondrial suspension (20-50 microg protein) is added to 1 mL assay buffer (50 mM KPi pH 7.5, 0.5 mM KCN, 100 microM DCIP) in a cuvette. Baseline is established, then NADH (100 microM) is added, and the linear decrease in absorbance at 600 nm is recorded. Rotenone (10 microM) is added as a control for Complex I-specific activity. All steps performed at 4degC except the assay (room temperature).
|
| ADME/Pharmacokinetics |
No pharmacokinetic data are available for DCIP hydrate sodium as it is not a drug. If administered in vivo, DCIP would likely be rapidly reduced to the colorless form by endogenous reductants (e.g., ascorbate, glutathione) and cleared from circulation. It is not absorbed intact from the gastrointestinal tract. The reduced form (DCIPH2) is highly unstable and would likely be rapidly oxidized or metabolized. No oral bioavailability, half-life, volume of distribution, or clearance values have been reported.
|
| Toxicity/Toxicokinetics |
DCIP hydrate sodium is an irritant and may cause skin and eye irritation. Based on its chemical structure (dichlorophenol derivative), there is potential for toxicity; the free phenol is known to be toxic. The compound is not classified as a carcinogen, mutagen, or reproductive toxin under GHS regulations. Acute oral LD50 has not been determined, but chlorinated phenols typically have LD50 values between 500-4000 mg/kg in rats. Avoid inhalation of powder and direct contact with skin. Use in fume hood with appropriate PPE (gloves, lab coat, safety goggles). In case of contact, flush with water.
|
| Additional Infomation |
DCIP hydrate sodium is not a drug and has no clinical or regulatory approval status. It is exclusively a laboratory reagent for research use. However, its derivative DPIP (the sodium salt) is used in educational laboratory experiments for photosynthesis measurement (Hill reaction) and vitamin C titration. The compound is also used in food science to detect reductants and preservatives. In pharmaceutical research, DCIP can be used to screen for inhibitors of Complex I in drug discovery programs targeting neurodegenerative diseases (e.g., Parkinson's disease), where mitochondrial dysfunction is implicated. No approved drugs are based on DCIP itself.
|
| Molecular Formula |
C12H8CL2NNAO3
|
|---|---|
| Molecular Weight |
308.0926
|
| Exact Mass |
306.977
|
| CAS # |
283164-79-4
|
| PubChem CID |
23676747
|
| Appearance |
Brown to black solid powder
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
19
|
| Complexity |
400
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1=CC(=CC=C1N=C2C=C(C(=O)C(=C2)Cl)Cl)[O-].O.[Na+]
|
| InChi Key |
XHSOLXWCGCVQHE-UHFFFAOYSA-M
|
| InChi Code |
InChI=1S/C12H7Cl2NO2.Na.H2O/c13-10-5-8(6-11(14)12(10)17)15-7-1-3-9(16)4-2-7;;/h1-6,16H;;1H2/q;+1;/p-1
|
| Chemical Name |
sodium;4-[(3,5-dichloro-4-oxocyclohexa-2,5-dien-1-ylidene)amino]phenolate;hydrate
|
| 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) |
DMSO: 100 mg/mL (324.58 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (8.11 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2458 mL | 16.2290 mL | 32.4580 mL | |
| 5 mM | 0.6492 mL | 3.2458 mL | 6.4916 mL | |
| 10 mM | 0.3246 mL | 1.6229 mL | 3.2458 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
