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| Targets |
Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 is not a pharmaceutical drug and does not target a biological receptor. It is a chemical labeling agent that functions as a ruthenium-based dye. The NHS ester reactive group specifically targets primary amine groups (-NH2) on proteins, peptides, and other amine-containing biomolecules. The ruthenium complex moiety serves as a sensitive electrochemiluminescence (ECL) label for detection and as a fluorescence quencher for quantum dots. It is also used as a capture probe for viral antigens.
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| ln Vitro |
Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 can be employed as a marker to determine the hydrodynamic radius of intravitreal anti-VEGF agents [2].
In vitro, Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 functions as a potent ruthenium-based dye for various bioanalytical applications. It is an effective quencher of quantum dot (QD) fluorescence, making it useful in FRET-based assays. It also serves as a capture probe for the virus antigen EV71, enabling the detection of this virus in diagnostic assays. Additionally, it functions as a sensitive electrogenerated chemiluminescence (ECL) label for the detection of matrix metalloproteinases (MMPs), with high sensitivity and selectivity. It has also been used as a labeling agent for measuring the hydrodynamic radii of intravitreal anti-VEGF therapeutics, demonstrating its utility in characterizing biopharmaceuticals. |
| ln Vivo |
In vivo, this compound is not typically administered directly, as it is a chemical labeling and detection reagent. However, it can be used to label biomolecules (e.g., antibodies, peptides) for use as imaging or diagnostic probes. A labeled antibody, for example, could be administered to an animal model to detect a specific antigen (e.g., tumor marker, virus antigen) using ECL or fluorescence detection methods. The ruthenium label is stable and provides a strong signal for sensitive in vitro assays of biological samples (e.g., serum, tissue homogenates) rather than for live animal imaging.
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| Enzyme Assay |
For non-cell-based ECL detection assays, a standard protocol for matrix metalloproteinase (MMP) detection uses a sandwich immunoassay format. A capture antibody specific for MMP is immobilized on a gold electrode or magnetic bead. The MMP protein is captured from the sample (e.g., cell culture supernatant or serum). A detection antibody conjugated to Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 is then added. After washing to remove unbound labeled antibody, the ECL signal is generated by applying a voltage in the presence of tripropylamine (TPA) as a co-reactant. The ECL intensity is measured using an electrochemiluminescence detector and is proportional to the concentration of MMP. The NHS ester group on the Ru complex can be used to conjugate the dye to any protein with free amine groups by incubating the protein (2-10 mg/mL) in PBS (pH 7.4-8.0) with a 10-20 fold molar excess of Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 for 1-2 hours at room temperature, followed by purification of the labeled conjugate.
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| Cell Assay |
For in vitro cell-based assays, Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 can be used to label antibodies for the detection of cell surface antigens. Cells (e.g., HeLa cells infected with EV71 virus) are fixed with 4% paraformaldehyde and blocked with 1% BSA. The Ru-labeled detection antibody (e.g., anti-EV71 antibody) is added and incubated for 60 minutes at room temperature. After washing, the cells are subjected to ECL detection on an ECL plate reader, or the Ru fluorescence can be detected using a fluorescence plate reader (excitation 458 nm, emission 628 nm). This approach enables high-sensitivity detection of viral antigens in infected cells. The compound can also be used as a capture probe for EV71 by immobilizing the Ru-labeled antibody on a solid surface for virus detection.
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| Animal Protocol |
This compound is not used directly in animal studies. Instead, it is used to label biomolecules (e.g., antibodies) for subsequent biodistribution or pharmacokinetic studies. For example, an antibody labeled with Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 can be administered intravenously to mice (10-20 mg/kg of labeled antibody). At various time points (1-72 hours), blood is collected, and organs are harvested. The concentration of the Ru-labeled antibody in plasma and tissue homogenates can be quantified by ECL or by measuring the Ru content using inductively coupled plasma mass spectrometry (ICP-MS). The labeled antibody can also be used in ex vivo imaging of tissue sections to visualize target antigen distribution.
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| ADME/Pharmacokinetics |
Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 has a molecular weight of 1014.66 g/mol and is supplied as a dark red to brown powder. It is soluble in organic solvents such as DMSO, DMF, and acetonitrile, and has moderate water solubility due to the presence of the charged PF6 counterions. The ruthenium complex is photostable and has a long fluorescence lifetime of near 500 nanoseconds, with large Stokes shifts and a lack of dye-dye interactions, making it suitable for time-resolved fluorescence assays. The NHS ester is prone to hydrolysis and should be used promptly after dissolution. For ECL applications, the compound exhibits high sensitivity and low background.
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| Toxicity/Toxicokinetics |
The compound contains ruthenium, a heavy metal, and hexafluorophosphate, which can be irritants. Hexafluorophosphate salts can hydrolyze to release hydrogen fluoride under certain conditions. Appropriate chemical safety precautions should be taken: use gloves, a lab coat, and eye protection. Avoid inhalation of dust. The compound should be stored at -20degC. No specific toxicology data is available, but heavy metal complexes can be cytotoxic at high concentrations. In ECL and labeling applications, the compound is used at low concentrations (nM to uM range) and is generally considered safe for research use. It is not intended for human or veterinary use.
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| References |
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| Additional Infomation |
Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 is a versatile ruthenium-based labeling reagent for bioanalytical applications. The NHS ester group provides a convenient method for conjugating the ruthenium complex to any amine-containing biomolecule, including proteins, antibodies, and peptides. The ruthenium complex exhibits a long fluorescence lifetime (near 500 ns), making it suitable for time-resolved fluorescence assays that eliminate background autofluorescence. The compound is also an effective electrochemiluminescence (ECL) label, with applications in clinical diagnostics (e.g., immunoassays for cancer biomarkers and infectious diseases). It is also used in FRET-based assays as a quencher of quantum dot fluorescence. This product is for research use only and is not intended for diagnostic or therapeutic applications.
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| Molecular Formula |
C36H29N7O4RU
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| Molecular Weight |
724.73000
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| Exact Mass |
725.132
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| CAS # |
136724-73-7
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| PubChem CID |
16218819
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| Appearance |
Brown to red solid powder
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| LogP |
5.897
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
24
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
62
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| Complexity |
662
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC(=NC=C1)C2=NC=CC(=C2)C(=O)ON3C(=O)CCC3=O.C1=CC=NC(=C1)C2=CC=CC=N2.C1=CC=NC(=C1)C2=CC=CC=N2.F[P-](F)(F)(F)(F)F.F[P-](F)(F)(F)(F)F.[Ru+2]
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| InChi Key |
UQJZJWDQEYXDCR-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H13N3O4.2C10H8N2.2F6P.Ru/c1-10-4-6-17-12(8-10)13-9-11(5-7-18-13)16(22)23-19-14(20)2-3-15(19)21;2*1-3-7-11-9(5-1)10-6-2-4-8-12-10;2*1-7(2,3,4,5)6;/h4-9H,2-3H2,1H3;2*1-8H;;;/q;;;2*-1;+2
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| Chemical Name |
(2,5-dioxopyrrolidin-1-yl) 2-(4-methylpyridin-2-yl)pyridine-4-carboxylate;2-pyridin-2-ylpyridine;ruthenium(2+);dihexafluorophosphate
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| HS Tariff Code |
2934.99.9001
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| 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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3798 mL | 6.8991 mL | 13.7982 mL | |
| 5 mM | 0.2760 mL | 1.3798 mL | 2.7596 mL | |
| 10 mM | 0.1380 mL | 0.6899 mL | 1.3798 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.
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