AO-EB double staining kit manual article number: CA1140
Specification: 100ml/500ml
Storage: 4°C protected from light and sealed, valid for one year Product introduction:
Acridine orange can penetrate into the intact cells of the cell membrane, insert into the nuclear DNA, and bind to the double-stranded DNA to emit green fluorescence. Ethidium bromide (EB) can only penetrate the damaged cells, embed nuclear DNA, and emit orange. Red fluorescence. The apoptotic cells appear to have enhanced staining, brighter fluorescence, uniform circular or condensed, agglomerate structures. Non-apoptotic nuclei exhibit structural features with varying fluorescence levels. Both are easy to judge. Under the fluorescence microscope, four cell morphology were observed: viable cells (VN), nuclear chromatin was green and showed normal structure; early apoptotic cells (VA), nuclear chromatin showed green pyknosis or beaded; Late apoptotic cells (NVA), nuclear chromatin is orange-red and condensed or beaded; non-apoptotic dead cells (NVN), nuclear chromatin orange-red and normal structure. The concentration of AO and EB solutions in this product is 100ug/ml, and the stabilizer contained does not affect the experimental results.
How to use: (for reference only)
Before use, the AO solution and the EB solution are mixed into a working solution according to the dosage, and the solution is used first.
For adherent cells or climbing slides, remove the medium, wash twice with PBS to remove residual medium and non-adherent cells, add new PBS; if you need to observe all cell characteristics, retain unattached cells, you can directly in the medium Add working fluid to it. Add 20 ul of working solution per ml of medium or PBS. After standing at room temperature for 2-5 min, it was observed under a fluorescence microscope.
For suspension cells, the working solution can be added directly to the working solution or centrifuged to collect the working solution in PBS. Add 20 ul of working solution per ml of medium or PBS. After standing at room temperature for 2-5 min, it was observed under a fluorescence microscope.
Precautions:
The phenol red-containing medium had a slight effect on the observation. A phenol-free red medium is recommended.
The working concentration and dyeing time of the dyeing solution can be adjusted according to the specific experimental conditions in order to achieve the best results.
Related reagents:
CA1020 ANNEXIN V-FITC/PI Apoptosis Detection Kit
CA1120 Hoechst 33342/PI Double Dye Kit
C0020 Hoechst 33258 staining solution (ready-to-use type)
C0060 DAPI solution (1mg/ml)
31800 RPMI Medium 1640 medium
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