Bitan-Lab wiki:Cell death by LDH assay

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cell death by LDH assay

Release of Lactate dehydrogenase through damage cell membrane indicate cell death which can be detected from culture media by CytoTox-ONETM Homogeneous Membrane Integrity Assay kit.

Reagents and accessories:

CytoTox-ONETM Homogeneous Membrane Integrity Assay kit (Promega), Black plate (Costar).

Protocol:

The cells were incubated with peptide for 48 h and cell death was assayed by measuring the release of lactate dehydrogenase (LDH) using according to the manufacturer’s instructions.

Briefly, 100 μl of substrate solution was added in each well and incubated for 20-30 min at room temperature.

After incubation 50 μl of stop solution was added and mix properly

The reading was taken at 340 nm using Synergy plate reader (Bio-TEK instruments, Winooski, VT)

Data were normalized with control and expressed as percentage cell death. The following formula was used to calculate the LDH (% cell death)

Cell viability (% of control) = {(OD of test sample-OD of full killed sample)/(OD of full killed sample-OD of control sample)}* 100

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