Ethidium Homodimer-III (EthD-III) is an alternative to Ethidium Homodimer-I (EthD-I). It exhibits a fluorescence spectrum similar to that of EthD-I; however, upon binding to DNA, EthD-III produces significantly brighter fluorescence than EthD-I. It is used for staining mammalian cells, bacteria, yeast, and fungi. EthD-III carries a strong positive charge, which prevents it from crossing cell membranes to stain live cells. Nevertheless, it enables accurate detection of nucleic acids in solution or in lysed cells, making it an exceptionally sensitive nucleic acid stain.Product Specifications1. Appearance: Red solid soluble in DMSO and MeOH2. λEx/λEm = 530/620 nm (when bound to DNA)3. λEx/λEm = 493 nm (when unbound to DNA)4. Molecular weight: ~1000ProtocolNote: This protocol is applicable to most cell types. Staining results may be affected by cell type, cell density, culture medium, and other factors; this protocol is for reference only.1. Stock solution preparation: Add an appropriate volume of DMSO to Ethidium Homodimer-I to prepare a 2 mM stock solution. This stock solution can be stably stored at -20°C for one year.2. Add 20 μL of the 2 mM stock solution to 10 mL of sterile, tissue-culture grade D-PBS, and mix thoroughly by vortexing to obtain a final concentration of 4 μM (recommended concentration range: 0.1–10 μM. For different cell lines, gradient optimization is recommended to determine the optimal staining concentration).3. Pipette 100–150 μL of the prepared working solution onto the cell coverslip to fully cover the cells. Incubation is preferably performed in a lidded dish to prevent evaporation of the staining solution.4. Incubate at room temperature for 30–45 minutes. Incubation time may be appropriately shortened if the staining solution concentration is too high or the temperature is too low.5. Add 10 μL of D-PBS to a new microscope slide.6. Using fine forceps, carefully and quickly invert the cell-bearing coverslip onto the slide containing D-PBS. To prevent evaporation of the staining solution, seal the edges of the coverslip with clear, transparent nail polish.7. Observe cell staining under a fluorescence microscope.

Specifications & Purity: Ethidium Homodimer-III (EthD-III) is an alternative to Ethidium Homodimer-I (EthD-I). It exhibits a fluorescence spectrum similar to that of EthD-I; however, upon binding to DNA, EthD-III produces significantly brighter fluorescence than EthD-I. It is used for staining mammalian cells, bacteria, yeast, and fungi. EthD-III carries a strong positive charge, which prevents it from crossing cell membranes to stain live cells. Nevertheless, it enables accurate detection of nucleic acids in solution or in lysed cells, making it an exceptionally sensitive nucleic acid stain.Product Specifications1. Appearance: Red solid soluble in DMSO and MeOH2. λEx/λEm = 530/620 nm (when bound to DNA)3. λEx/λEm = 493 nm (when unbound to DNA)4. Molecular weight: ~1000ProtocolNote: This protocol is applicable to most cell types. Staining results may be affected by cell type, cell density, culture medium, and other factors; this protocol is for reference only.1. Stock solution preparation: Add an appropriate volume of DMSO to Ethidium Homodimer-I to prepare a 2 mM stock solution. This stock solution can be stably stored at -20°C for one year.2. Add 20 μL of the 2 mM stock solution to 10 mL of sterile, tissue-culture grade D-PBS, and mix thoroughly by vortexing to obtain a final concentration of 4 μM (recommended concentration range: 0.1–10 μM. For different cell lines, gradient optimization is recommended to determine the optimal staining concentration).3. Pipette 100–150 μL of the prepared working solution onto the cell coverslip to fully cover the cells. Incubation is preferably performed in a lidded dish to prevent evaporation of the staining solution.4. Incubate at room temperature for 30–45 minutes. Incubation time may be appropriately shortened if the staining solution concentration is too high or the temperature is too low.5. Add 10 μL of D-PBS to a new microscope slide.6. Using fine forceps, carefully and quickly invert the cell-bearing coverslip onto the slide containing D-PBS. To prevent evaporation of the staining solution, seal the edges of the coverslip with clear, transparent nail polish.7. Observe cell staining under a fluorescence microscope.

Grade: BioReagent;Biological Stain;for Fluorescence analysis;for Microscopy

Molecular Formula: C51H62Cl5N9

Molecular Weight: 978.36

Storage Temperature: Protected from light;Store at -20°C;Argon charged

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