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| Targets |
The primary molecular target of LDS-751 is nucleic acids, specifically DNA, to which it binds with high affinity. The compound is a cell-permeant nucleic acid stain that intercalates or binds to DNA, resulting in fluorescence enhancement upon binding. The dye binds to double-stranded DNA, resulting in a fluorescence signal that is proportional to the DNA content of the cell. LDS-751 does not have a specific protein target; rather, it functions as a nucleic acid-binding fluorescent probe. Its mechanism of action involves the intercalation or minor groove binding to DNA, which restricts the rotation of the dye molecule and leads to increased fluorescence quantum yield. The dye's ability to penetrate cell membranes allows it to stain nucleic acids in intact cells without the need for cell permeabilization.
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| ln Vitro |
In vitro studies of LDS-751 focus on its fluorescent properties and its use as a nucleic acid stain. The compound exhibits high affinity for DNA and undergoes fluorescence enhancement upon binding, with a maximum emission wavelength of 670 nm. The dye can be well excited with a 488 nm laser line, although it has a peak excitation at approximately 543 nm on double-stranded DNA. LDS-751 is used in combination with Thiazole Orange for multiplexed staining applications. The compound's stability in various buffers and its behavior under different staining conditions have been characterized. It is a valuable tool for flow cytometry, fluorescence microscopy, and other fluorescence-based applications. The dye's fluorescence properties, including excitation and emission spectra, have been well characterized, enabling its use in a wide range of assay formats. In cellular studies, LDS-751 is used as a cell-permeant nucleic acid stain for flow cytometry and fluorescence microscopy applications. The compound can penetrate cell membranes and bind to nucleic acids within cells, enabling the visualization and quantification of DNA content. LDS-751 is used to distinguish intact nucleated cells from anucleated and damaged nucleated cells, and to distinguish different cell types in mixed populations of neutrophils, lymphocytes, and monocytes by flow cytometry. In flow cytometry, LDS-751 staining allows for the identification and sorting of cell populations based on their nucleic acid content. The compound's fluorescence is enhanced upon binding to DNA, providing bright and specific staining. LDS-751 is also used in combination with other fluorescent probes for multiplexed analysis of cellular parameters. Its cell-permeability and DNA-binding properties make it a useful tool for cell biology research.
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| ln Vivo |
In vivo studies of LDS-751 are limited, as the compound is primarily used as a research reagent for in vitro and cellular applications rather than as a therapeutic or diagnostic agent. However, fluorescent probes like LDS-751 may be used in animal models for imaging applications, such as tracking cells or biomolecules in vivo. In such studies, LDS-751-labeled cells or molecules are administered to animals, and their distribution and fate are monitored by fluorescence imaging. The compound's fluorescence properties enable sensitive detection in biological samples. No extensive in vivo pharmacological or toxicological studies have been reported for LDS-751.
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| Enzyme Assay |
For fluorescence characterization assays, LDS-751 is dissolved in appropriate solvent (e.g., DMSO, methanol, or aqueous buffer) and its absorption and emission spectra are measured using a UV-Vis spectrophotometer and fluorescence spectrometer, respectively. Excitation and emission maxima are determined. For nucleic acid binding studies, LDS-751 is incubated with purified DNA in assay buffer (e.g., 10 mM Tris-HCl, pH 7.4, 100 mM NaCl) at varying concentrations (0.1-10 µM). Fluorescence intensity is measured at the emission maximum, and the fluorescence enhancement upon DNA binding is calculated. Binding affinity is determined by fitting fluorescence titration data to a binding isotherm. For flow cytometry applications, the optimal staining concentration and incubation time are determined by titrating the dye in cells and measuring the fluorescence signal and signal-to-noise ratio.
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| Cell Assay |
For cellular staining, cells (e.g., whole blood, cultured cells, or isolated leukocytes) are harvested and resuspended in PBS or appropriate buffer at a density of 1-5 × 10⁶ cells/ml. LDS-751 is added to the cell suspension at a final concentration of 0.1-10 µM (typically 1-5 µM) and incubated at room temperature or 37°C for 15-30 minutes in the dark. Stained cells are washed twice with PBS to remove unbound dye. For flow cytometry analysis, stained cells are analyzed using a flow cytometer equipped with an appropriate laser line (e.g., 488 nm) and fluorescence detectors. Fluorescence intensity is measured in the appropriate channel (e.g., FL2 or FL3, typically 670 nm emission). For fluorescence microscopy, stained cells are placed on glass slides or chamber slides and imaged using a fluorescence microscope with appropriate filter sets. For combination staining with other dyes (e.g., Thiazole Orange), cells are stained with both dyes simultaneously or sequentially, and fluorescence signals are measured in different channels.
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| Animal Protocol |
No standard in vivo protocols exist specifically for LDS-751, as it is primarily a research reagent for in vitro and cellular applications. If used in animal studies, LDS-751 would likely be administered intravenously or intraperitoneally to mice at doses determined from preliminary studies. Tissues and organs would be collected at various time points and processed for fluorescence imaging or homogenized for fluorometric analysis. Biodistribution would be assessed by measuring fluorescence in tissue homogenates or by imaging whole animals using an in vivo imaging system. However, such studies are not typical for this compound, and no specific dosing or administration protocols are reported in the literature.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for LDS-751 are not available, as the compound is used as a research reagent rather than a therapeutic agent. If used in vivo, LDS-751 would be expected to have distribution to various tissues due to its cell-permeability. The compound's metabolism and excretion would depend on its chemical structure, likely involving hepatic metabolism and biliary or renal clearance. However, no dedicated ADME studies have been reported for LDS-751. The compound is primarily used in vitro and is not intended for in vivo pharmacokinetic characterization.
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| Toxicity/Toxicokinetics |
Toxicological data for LDS-751 are limited, as the compound is used as a research reagent and not as a therapeutic agent. The compound is generally considered to have low toxicity at the concentrations used for staining applications (typically 0.1-10 µM). No acute toxicity, mutagenicity, or carcinogenicity data have been reported. As with all chemical reagents, appropriate safety precautions should be taken when handling LDS-751, including the use of personal protective equipment (gloves, lab coat, safety glasses) and work in a well-ventilated area. The compound should be handled according to standard laboratory safety protocols.
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| References |
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| Additional Infomation |
LDS 751 dye is an organic perchlorate. It is used as a fluorescent dye. It contains LDS 751(1+).
LDS-751 (CAS 181885-68-7) is a fluorescent, cell-permeant nucleic acid stain that primarily detects DNA. It is widely employed in flow cytometry for staining nucleic acids within cells, allowing researchers to distinguish between intact nucleated cells and non-nucleated or damaged cells. LDS-751 has high affinity for DNA and undergoes fluorescence enhancement upon binding, with maximum emission at 670 nm. It can be excited with a 488 nm laser line, with peak excitation at ~543 nm on dsDNA. The dye is used in combination with Thiazole Orange for multiplexed applications. Its molecular formula is C₂₅H₃₀N₃·ClO₄ with a molecular weight of 471.98. LDS-751 is a valuable tool for cell biology and flow cytometry research and is strictly for research use only. |
| Molecular Formula |
C25H30N3.O4CL-
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| Molecular Weight |
471.9764
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| Exact Mass |
471.192
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| CAS # |
181885-68-7
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| PubChem CID |
5706752
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| Appearance |
Green to black solid powder
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
33
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| Complexity |
612
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC[N+]1=C(C=CC2=C1C=CC(=C2)N(C)C)/C=C/C=C/C3=CC=C(C=C3)N(C)C.[O-]Cl(=O)(=O)=O
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| InChi Key |
FGBAVQUHSKYMTC-UHFFFAOYSA-M
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| InChi Code |
InChI=1S/C25H30N3.ClHO4/c1-6-28-23(16-13-21-19-24(27(4)5)17-18-25(21)28)10-8-7-9-20-11-14-22(15-12-20)26(2)3;2-1(3,4)5/h7-19H,6H2,1-5H3;(H,2,3,4,5)/q+1;/p-1
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| Chemical Name |
2-[(1E,3E)-4-[4-(dimethylamino)phenyl]buta-1,3-dienyl]-1-ethyl-N,N-dimethylquinolin-1-ium-6-amine;perchlorate
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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: (1). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. (2). This product is not stable in solution, please use freshly prepared working solution for optimal results. |
| 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) |
DMSO : ~83.33 mg/mL (~176.55 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.41 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.41 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 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 | 2.1187 mL | 10.5937 mL | 21.1873 mL | |
| 5 mM | 0.4237 mL | 2.1187 mL | 4.2375 mL | |
| 10 mM | 0.2119 mL | 1.0594 mL | 2.1187 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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