MitoP, IC-03915003
Product Introduction
MitoP is the product of the stoichiometric reaction between the mitochondria-targeted hydrogen peroxide (H₂O₂) probe MitoB and mitochondrial H₂O₂. MitoB, consisting of a triphenylphosphonium (TPP) lipophilic cation conjugated to an arylboronic acid moiety, selectively accumulates in the mitochondrial matrix and reacts specifically with H₂O₂ to yield MitoP. As the direct reaction product, MitoP serves as an essential reference standard for the quantitative mass spectrometry-based measurement of mitochondrial H₂O₂ levels using the MitoB/MitoP probe system. Accurate quantification of the MitoP/MitoB ratio by LC-MS/MS requires a purified MitoP standard for calibration, enabling absolute determination of MitoP concentrations in biological samples. MitoP is also valuable for studies investigating the mitochondrial import, accumulation, and metabolism of TPP-conjugated compounds independent of the H₂O₂-sensing chemistry. This product provides a high-purity reference standard indispensable for researchers employing the MitoB probe for in vitro and in vivo mitochondrial H₂O₂ measurements.
Product Features
1. MitoB Reaction Product Standard: Essential reference compound for the quantitative LC-MS/MS analysis of mitochondrial H₂O₂ using the MitoB/MitoP ratiometric probe system.
2. Analytical Calibration Standard: Enables accurate quantification of MitoP in biological samples by providing a purified reference for calibration curve generation.
3. TPP Moiety for Mitochondrial Targeting: Contains the triphenylphosphonium cation, facilitating studies of mitochondrial import and accumulation of TPP-conjugated compounds.
4. High Purity: Supplied at high purity suitable for use as an analytical reference standard in mass spectrometry applications.
5. Complementary to MitoB Probe: Designed for use alongside MitoB (IC-03914995) for complete mitochondrial H₂O₂ measurement workflows.
Specifications
Size: Available in 1 mg and 5 mg
CAS Number: 74597-01-6
Molecular Weight: 458.50
Molecular Formula: C30H34O2P
Purity: ¡Ý98%
Solubility: DMSO: ¡Ý20 mg/mL; Ethanol: ¡Ý10 mg/mL; limited solubility in aqueous buffers
Storage and Stability
Storage Conditions: Store at -20¡ãC, protected from light and moisture.
Shelf Life: The product is stable for 24 months from the date of manufacture when stored as directed. For reconstituted solutions, aliquot and store at -80¡ãC. Avoid repeated freeze-thaw cycles.
Protocol (For Reference Only)
Important: MitoP is intended as an analytical reference standard for LC-MS/MS calibration. Protect from light and moisture. Prepare stock solutions freshly before use.
1. Stock Solution Preparation: Dissolve MitoP in anhydrous DMSO or ethanol to prepare a concentrated stock solution (e.g., 10-100 mM). Aliquot into single-use vials and store at -80¡ãC protected from light.
2. Calibration Standard Preparation: For LC-MS/MS calibration, prepare a serial dilution of MitoP in extraction solvent (typically 60% acetonitrile in water containing 0.1% formic acid) to generate a calibration curve spanning the expected concentration range in biological samples (typically 0.1-1000 nM). Include an appropriate internal standard as recommended for the specific analytical method.
3. Sample Processing for MitoB/MitoP Ratio Analysis: When used in conjunction with MitoB-treated biological samples, homogenize cell pellets or tissue samples in ice-cold extraction solvent spiked with a known concentration of internal standard. Centrifuge at 16,000 x g for 10 minutes at 4¡ãC to remove precipitated protein. Analyze the supernatant by LC-MS/MS alongside the MitoP calibration standards.
4. LC-MS/MS Analysis: Analyze MitoP using positive electrospray ionization with multiple reaction monitoring (MRM) transitions optimized on your instrument. Typical MRM parameters can be established by direct infusion of the MitoP standard. Quantify MitoP in unknown samples by interpolation from the calibration curve.
5. Data Normalization: Calculate the MitoP/MitoB ratio for each sample. The ratio is proportional to the average mitochondrial H₂O₂ concentration integrated over the MitoB incubation period. Use vehicle-treated controls to determine basal H₂O₂ levels.
Precautions
1. MitoP is an analytical reference standard; it is not a probe and does not detect H₂O₂. It is the chemically defined product of the MitoB-H₂O₂ reaction used for calibration.
2. Protect from light and moisture during storage; prepare stock solutions in anhydrous solvents.
3. For LC-MS/MS quantification, optimize instrument parameters for your specific system using direct infusion of MitoP standard.
4. For in vivo MitoB studies, tissue harvesting and processing must be performed rapidly to prevent post-mortem changes that may alter the MitoP/MitoB ratio.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What is the difference between MitoB and MitoP?
A1: MitoB is the mitochondria-targeted H₂O₂ probe containing an arylboronic acid reactive group. Upon reaction with mitochondrial H₂O₂, MitoB is stoichiometrically converted to MitoP. MitoB is administered to cells or animals for H₂O₂ detection; MitoP is the reference standard used for calibrating the LC-MS/MS measurement of the MitoB-to-MitoP conversion.
Q2: Do I need MitoP if I am using MitoB for mitochondrial H₂O₂ measurement?
A2: Yes. Accurate quantification of the MitoP/MitoB ratio by LC-MS/MS requires a purified MitoP standard for calibration curve generation. Without the MitoP standard, absolute quantification of MitoP in biological samples is not possible, and results can only be expressed as relative ratios.
Q3: Can MitoP be used independently of MitoB?
A3: MitoP is designed specifically as the analytical standard for MitoB-based H₂O₂ measurements. It may also be used to study the mitochondrial accumulation and metabolism of TPP-conjugated compounds independently of the H₂O₂-sensing chemistry.
Q4: What mass spectrometry parameters should I use for MitoP detection?
A4: MitoP is typically analyzed by LC-MS/MS using positive electrospray ionization with MRM transitions. The specific transitions depend on your instrument configuration. Optimize parameters by direct infusion of MitoP standard in the presence of mobile phase components.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of analytical parameters and sample preparation is recommended for specific experimental systems.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: IC-03915003
Product Name: MitoP
Size: 1 mg / 5 mg
Price:
1 mg: CNY ¥1050.00 / USD $105.00 / EUR €126.00 / JPY ¥18900.00
5 mg: CNY ¥2850.00 / USD $285.00 / EUR €342.00 / JPY ¥51300.00
2023 Version
