Live-Dead Cell Staining Kit

(Catalog #K501-100; 100 stainings; Store kit at –20^oC)

I. Introduction:

Distinguishing between live and dead cells is very important for investigation of growth control and cell death. The Live-Dead Cell Staining Kit provides the ready-to-use reagents for convenient discrimination between live and dead cells. The kit utilizes Live-Dye^TM, a cell-permeable green fluorescent dye (Ex/Em = 488/518 nm), to stain live cells. Dead cells can be easily stained by propidium iodide (PI), a cell non-permeable red fluorescent dye (Ex/Em = 488/615). Stained live and dead cells can be visualized by fluorescence microscopy using a band-pass filter (detects FITC and rhodamine). The kit provides sufficient reagents for 100 stainings using 24-well plate.

 

1. 
   

   Kit Contents:

   
   

 

 

 

 

2. 
   

   Cell Staining Protocol:

   
   

1.. Prepare enough Staining Solution for your assay (0.5 ml per well in 24 well dish): mix 1 μl of Solution A and 1 μl of Solution B in 1 ml of Staining Buffer. Scale up accordingly for larger numbers of assays.

1. 
   

   Collect cells (1 x 10⁶ cells) by centrifugation at 500 X g for 5 minutes.

   
   


2. 
   

   Resuspend to 0.5 ml Staining Solution

   
   


3. 
   

   Incubate for 15 minutes at 37^oC.

   
   


4. 
   

   Place the cell suspension on a glass slide. Cover the cells with a glass coverslip.

   
   

For analyzing adherent cells, grow cells directly on a coverslip. Following incubation with the Staining Solution, invert coverslip on a glass slide and visualize cells.

5. 
   

   Observe cells immediately under a fluorescence microscope using a band-pass filter (detects fluorescein and rhodamine).

   
   

Healthy cells stain only the cell-permeable Live-Dye, fluorescing green. Dead cells can stain both the cell-permeable Live-Dye and the cell non-permeable PI (red), the overlay of green and red appears to be yellow-red.

3. 
   

   Caution:

   
   

As the optimal staining conditions may vary among different cell types, we recommend that a suitable concentration of Solution A and B be determined individually.

Please note that PI is suspected to be highly carcinogenic, so careful handling of the reagent is required.

 

V. Storage and Stability:

Store kit at –20^oC. Protect from light. Store Staining Buffer at 4^oC after opening. All reagents are stable for 1 year under proper storage conditions.