Live/Dead cell Staining Kit II

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Assay performed atdifferent time intervals (24, 48, and 72 h)	Add Calcein-AM/EthD-III staining solution and incubate the cells for 30-45 minutes at room temperature.

Upstream tips
Assay performed atdifferent time intervals (24, 48, and 72 h)
Protocol tips
Add Calcein-AM/EthD-III staining solution and incubate the cells for 30-45 minutes at room temperature.

Publication protocol

n vitro iPSC assays for cell viability (n=3) (live/dead cell staining kit II, Promokine; PromoCell) and proliferation (n=3) (WST-8 assay, Promokine; PromoCell) were performed on 3 consecutive days. To differentiate living from dead cells, 3×104 iPSCs were seeded on a 24-well plate. Well chambers were assigned to three groups: fibrinogen, fibrin, and medium. In each group, chambers were assigned for the respective time point at which measurements were performed: 24, 48, and 72 h. At each time point, cells were stained: green, living cells; red, dead cells. Following microscopy, representative areas of live and dead cells were measured using ImageJ (v. 1.48 for Mac OS X).

In vitro cell proliferation analysis was performed applying the WST-8 assay (Promokine; PromoCell). A 96-well plate was arranged for cell culture with three different groups: fibrin, fibrinogen and medium. Every group also included noncellular controls. In each well chamber 5×103 iPSCs were seeded and brought into culture. Separately, standard dilutions of iPSCs ranging from 2.5×103 to 1×104 cells were assembled at each time point for cell quantification. Quantification was performed using a microplate reader (Paradigm Detection Platform; Beckman Coulter) at 24, 48, and 72 h applying the WST-8 substrate.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Live / Dead assay mammalian cells - mouse iPSC using Live/Dead cell Staining Kit II from PromoKine.

Manufacturer protocol

Download the product protocol from PromoKine for Live/Dead cell Staining Kit II below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Live / Dead assay mammalian cells - mouse iPSC using Live/Dead cell Staining Kit II from PromoKine. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.