Annexin V-EGFP/PI Apoptosis Assay Kit

The Annexin V-EGFP/PI Apoptosis Assay Kit is designed to detect early apoptosis using EGFP-labeled Annexin V. This molecular probe detects the presence of phosphatidylserine (PS) on the cell membrane surface, which is a characteristic of early apoptotic cells. Normally, PS is located on the inner side of the cell membrane, but when the cell undergoes apoptosis, PS flips to the outer surface of the membrane and is exposed to the extracellular environment. Annexin - Ⅴ is a protein with high affinity for PS, and can bind to the membrane of early apoptotic cells. To distinguish surviving early cells from necrotic or late apoptotic cells, the kit provides Propidium Iodide (PI), a nucleic acid dye that can cross the membrane boundary of late apoptotic and necrotic cells to make the nucleus red. By combining Annexin V with PI, the kit can exclude living cells (Annexin V-/PI-) and early apoptotic cells (Annexin V+/PI-) while identifying late apoptotic and necrotic cells (Annexin V+/PI+). Using this kit, researchers can detect early apoptosis and differentiate it from other forms of cell death.
Product components:
- Code name: 20T
- Code name: 50T
- Component A: Annexin V-EGFP, available in 100ul and 250ul volumes
- Component B: PI, available in 100ul and 250ul volumes
- Component C: Binding Buffer, available in 5ml and 10ml volumes.
Please note that the generated content above is based on the original text information provided.

1×10^6/ml。，。
To start with, carefully move 0.2 ml of the cell suspension from the cell culture flask containing 2×10^5 cells and transfer it to a centrifuge tube that has been properly cleaned.
1000 g5，。
The cells that had come out of precipitation were carefully mixed with 0.2 ml of chilled PBS solution to resuspend them.
Begin centrifugation for a duration of 5 minutes at room temperature and at a speed of 1000 g. Once completed, remove the PBS solution from the sample.
The cell precipitation was resuspended by adding 0.2ml of precooled Binding Buffer (component C, also known as Binding Buffer). The resuspension was done gently to ensure proper mixing of the cells with the buffer.
First, gently mix 5 μl of Annexin V-EGFP (Component A) with 5 μl of PI (Component B).
Leave the mixture to incubate in darkness at room temperature (18-24°C) for a period of 10-15 minutes.
If desired, additional buffer or PBS may be added to bring the volume up to 0.4ml. To create similar content, the text can be rephrased as follows: To increase the overall volume, it is possible to supplement the mixture with a binding buffer or PBS until a final volume of 0.4ml is achieved.
As soon as possible, place the sample on the ice and ensure it is kept away from light. It is imperative to test the sample promptly.
Detection and analysis were conducted using flow cytometry or fluorescence microscopy.

Store at 2-8 ° C away from light for 6 months.
For optimal storage, it is recommended to keep the product in a dark location at a temperature of -20 ° C or below. Care should also be taken to prevent repeated freeze-thaw cycles, as this could compromise the quality of the product over time. With proper storage, the product should remain viable for at least 12 months, if not longer.

Annexin V-EGFP/PI Apoptosis Detection Kit