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Description
AO/PI Double StainingProduct Specification Usage 1. Reagent preparation: (1) PBS buffer (pH7. 4) (2) or HbSS (hank's balanced salt solution) 2. Consumables preparation (1) centrifuge tube (2) suction tip (3) disposable gloves 3. Dyeing working solution preparation: (1) according to the number of samples, prepare the staining buffer according to the following ratio. Take 100 L reagent C used 900 Dilute with L pure water and mix well to form staining buffer (2) every 500
Product Specification
| Usage | 1. Reagent preparation: (1) PBS buffer (pH7.4) (2) or HbSS (hank's balanced salt solution) 2. Consumables preparation (1) centrifuge tube (2) suction tip (3) disposable gloves 3. Dyeing working solution preparation: (1) according to the number of samples, prepare the staining buffer according to the following ratio. Take 100μ L reagent C used 900μ Dilute with L pure water and mix well to form staining buffer (2) every 500 mu; L staining buffer, add 5ul Ao staining solution and 10ul PI staining solution, and mix well to form the staining working solution 4. Suspension cell staining (1) collect sample cells, and the number of cells is within 10x105 (2) wash the cells twice with PBS (3) use 500 mu; L staining working solution to resuspend the cells (4) incubate for 10-20 minutes at 4 ℃ in the dark after gently mixing (5) wash the cells with PBS (6) the results were detected by fluorescence microscope or flow cytometry 5. In situ staining of adherent cells (1) wash the cells twice with PBS (2) add an appropriate volume of staining working solution to the cell culture plate or cell climbing piece (3) incubate the cells at 37 ℃ for 10-20 minutes, and the optimal culture time is different for different cells. 20 min can be used as the initial incubation time, after which the system can be optimized to obtain uniform labeling results (4) blot the staining working solution, wash the culture plate or coverslip with medium for 2-3 times, cover all cells with preheated medium each time, and then blot the medium Result Analysis: Under the fluorescence microscope, 488nm excitation light was used for microscopic examination: the DNA of normal cells stained with AO was uniform yellow or yellow green, and the morphology and structure were normal when cells are apoptotic, chromatin condenses, and nuclei are fragmented into punctate, which are stained in a dense and dense manner with different sizes Necrotic cells showed strong red fluorescence |
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| Description |
AO/PI double staining apoptosis detection kit is a commonly used staining method for apoptosis morphology research, which uses AO/PI probe to double stain the nucleus to detect the state of apoptotic cells
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| General Notes | 1. The reagent in the screw cap micro reagent tube should be centrifuged briefly before opening the cap, and the liquid on the inner wall of the cap should be collected to the bottom of the tube to avoid liquid spilling when opening the cap 2. Careful operation is required for cell treatment to avoid artificial damage to cells 3. This staining kit can be used for cells, cell smears, etc. The following is an example of fluorescence microscopy detection of suspension cultured cells. For other methods, please refer to relevant materials 4. Adherent cells can be digested and stained, or directly stained in situ. |
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| Storage Temp. | 2-8 ℃ protected from light, 6months | ||||||||||||||||
| Applications | Flow cytometry; Laser confocal; fluorescence microscope |
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