K2081-1000T | Live-Dead Cell Staining Kit Clinisciences

Product Citation

1. Yeyang Wang, et al. "Electrospun silk fibroin/fibrin vascular scaffold with superior mechanical properties and biocompatibility for applications in tissue engineering." Sci Rep. 2024 Feb 16;14(1):3942. PMID: 38365964
2. Junyu Chen, et al. "Magnetic scaffold constructing by micro-injection for bone tissue engineering under static magnetic field." Journal of Materials Research and Technology. Volume 29, March–April 2024, Pages 3554-3565.
3. Tanchen Ren, et al. "Enhancing aortic valve drug delivery with PAR2-targeting magnetic nano-cargoes for calcification alleviation." Nat Commun. 2024 Jan 16;15(1):557. PMID: 38228638
4. Chao Zhang, Hong Huang, et al. "DNA supramolecular hydrogel-enabled sustained delivery of metformin for relieving osteoarthritis." ACS Appl Mater Interfaces. 2023 Apr 5;15(13):16369-16379. PMID: 36945078

Description

Our product Live-Dead Cell Staining Kit is used for simultaneous fluorescence staining of viable and dead cells. This kit contains two dyes-Calcein-AM and Propidium Iodide (PI), which stain viable and dead cells, respectively.

Calcein-AM, acetoxymethyl ester of calcein, is highly lipophilic and cell membrane permeable. Though Calcein-AM itself isn’t a fluorescent molecule, it will turn to Calcein after esterase’s digestion in a viable cell, which could emit strong green fluorescence (excitation: 490 nm, emission: 515 nm). Therefore, Calcein-AM only stains viable cells.

In contrast, PI, a nuclei staining dye, can’t pass through a viable cell membrane. It reaches the nucleus by passing through disorderedareas of dead cell membrane, and intercalates with the DNA double helix of the cell to emit red fluorescence (excitation: 535 nm, emmision: 617 nm).

Since both calcein and PI-DNA can be excited with 490 nm, simultaneous monitoring of viable and dead cells is possible with a fluorescence microscope or flow cytometer. With 545 nm excitation, only dead cells can emit red fluorescence. Since optimal staining conditions constantly changes because of different cell lines, we recommend that a suitable concentration of PI and Calcein-AM should be individually determined.

Please note that PI is suspected to be highly carcinogenic; careful handling is required.

Quality Control

Protocol

Components and Storage

	500 Tests	1000 Tests
Calcein-AM Solution (2 mM)	50 μL	50 μL × 2
PI Solution (1.5 mM)	150 μL	150 μL × 2
Store at -20°C, protect from light. Calcein-AM is easily hydrolized by moisture. Keep it desiccate and tightly close the cap after the use.

Storage

Store at -20°C, protect from light. Calcein-AM is easily hydrolized by moisture. Keep it desiccate and tightly close the cap after the use.

Live-Dead Cell Staining Kit

Referentie K2081-1000T

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