Live / Dead assay mammalian cells - rat nucleus pulposus

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

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Found 2 matching solutions for this experiment

Cell Counting Kit-8

Dojindo

Upstream tips	Protocol tips	Downstream tips
	Add 10 μl CCK-8 solution incubate for 2 h.	Absorbance of each wells was measured at 450 nm To measure the absorbance later, add 10 μl of 1% w/v SDS or 0.1 M HCl to each well, cover the plate and store it with protection from light at room temperature. No absorbance change should be observed for 24 hours

Protocol tips
Add 10 μl CCK-8 solution incubate for 2 h.
Downstream tips
Absorbance of each wells was measured at 450 nm To measure the absorbance later, add 10 μl of 1% w/v SDS or 0.1 M HCl to each well, cover the plate and store it with protection from light at room temperature. No absorbance change should be observed for 24 hours

Manufacturer protocol Publication protocol 1 Relevant paper

LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
Wash cells 2–3 times with PBS	Add calcein AM: 2 μM; EthD-1: 4 μM Incubate the cells for 40 minutes at room temperature

Upstream tips
Wash cells 2–3 times with PBS
Protocol tips
Add calcein AM: 2 μM; EthD-1: 4 μM Incubate the cells for 40 minutes at room temperature

Manufacturer protocol Publication protocol 1 Relevant paper

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