Live-Dead Cell Staining Kit活死細(xì)胞染色試劑盒
(Catalog #K501-100; 100 stainings; Store kit at –20°C)
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-
DyeTM, 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.
II. Kit Contents:Component K501-100 Part Number
Solution A (1 mM Live-Dye) 50 μl K501-100-1
Solution B (2.5 mg/ml PI) 50 μl K501-100-2
Staining Buffer 50 ml K501-100-3
III. 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.
2. Collect cells (1 x 106cells) by centrifugation at 500 X g for 5 min.
3. Resuspend to 0.5 ml Staining Solution
4. Incubate for 15 min at 37°C.
5. 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.
6. Observe cells immediay 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.
IV. 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°C. Protect from light. Store Staining Buffer at 4°C after opening. All
reagents are stable for 1 year under proper storage conditions.
FOR RESEARCH USE ONLY! Not to be used on humans.
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