Solarbio's Hoechst 33342 /PI double staining kit provides you with a classic, fast and easy method for the detection of apoptosis and cell necrosis. This kit was double-stained with Hoechst 33342 and Propidium Iodide (PI). When cells undergo apoptosis, chromatin shrinks. Hoechst33342 can penetrate the cell membrane, and the fluorescence of apoptotic cells will be significantly enhanced than that of normal cells after staining. Propyridine iodide (PI) cannot penetrate cell membranes and cannot stain normal or apoptotic cells with intact cell membranes. For necrotic cells, the integrity of their cell membranes is lost, and propyridine iodide (PI) can stain necrotic cells. When the two dyes were double-dyed, the normal cells showed weak red fluorescence + weak blue fluorescence, the apoptotic cells showed weak red fluorescence + strong blue fluorescence, and the necrotic cells showed strong red fluorescence + strong blue fluorescence. Referring to the following figure, the left picture shows normal cells before inducing apoptosis, and the right picture shows cells after inducing apoptosis.

100T

500T

细胞染色缓冲液

100ml

500ml

Hoechst染色液

0.5ml

2.5ml

PI染色液

0.5ml

2.5ml

说明书

1份

Instruction:

1. About 100 to 1 million cells were collected for each sample in a 1.5ml centrifuge tube, and the supernatant was centrifuged. Cell precipitates were resuspended with 0.8-1ml cell stain buffer.

2. Add 5 μl Hoechst staining solution.

3. Add 5 microliters of PI dyeing solution.

4. Mix well and incubate in ice bath or at 4℃ for 20-30 minutes.

5. Red fluorescence and blue fluorescence were detected by flow cytometry.

6. If fluorescence microscopy is used, centrifuge the precipitated cells before detection, wash them once with PBS, and then observe the red fluorescence and blue fluorescence with smear. For adherent cells, fluorescence microscopy can be used to detect cells without collecting cells, and directly add the cell staining buffer, Hoechst staining solution and PI staining solution to ice bath or 4℃ for 20-30 minutes according to the above proportion. After staining, wash with PBS once and observe under fluorescence microscope.

Note:

Flow cytometry is required for red and blue dual fluorescence detection. Fluorescence microscopy can also be used. Test as soon as possible after staining.

Hoechst 33342 is harmful to human body, propyl iodide (PI) is irritating to human body, please pay attention to appropriate protection.

For your safety and health, please wear a lab coat and disposable gloves.

Note：Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.