LDH-Cytotoxicity Colorimetric Assay Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Add 100 µl LDH Reaction Mix to each well, mix and incubate for 30 min at room temperature- - The plate can be read at multiple time points until the desired reading is observed. The high control should be OD450nm ~ 2.0, while the low control should be OD450nm < 0.8.

Protocol tips
- Add 100 µl LDH Reaction Mix to each well, mix and incubate for 30 min at room temperature- - The plate can be read at multiple time points until the desired reading is observed. The high control should be OD450nm ~ 2.0, while the low control should be OD450nm < 0.8.

Publication protocol

The LDH release assay was used as an index of cell toxicity and was done as described previously 31 with minor modifications. In this assay, LDH activity in the cultured medium represented LDH release from SH‐SY5Y cells treated with ionomycin or vehicle (DMSO only) for the indicated times in the absence or presence of TSA pretreatment. In brief, cells cultured in 12‐well plates were pretreated with 300 nm TSA for 24 h and thereafter co‐treated further with 5 μm ionomycin for the indicated times. For low control, cells were cultured without treatment. A high control was prepared by treating cells with 1% Triton X‐100. Test samples were prepared by treating cells with DMSO, TSA, ionomycin or TSA plus ionomycin. An aliquot (100 μL) of the cultured medium was collected into a 96‐well plate and incubated with 100 μL of reaction mixture, and the LDH release assay was performed using an LDH cytotoxicity assay kit (BioVision, Mountain View, CA, USA), according to the manufacturer's instructions. Cytotoxicity was calculated using the equation cytotoxicity (%) = [(test sample − low control)/(high control − low control)] × 100.

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Manufacturer protocol

Download the product protocol from Biovision for LDH-Cytotoxicity Colorimetric Assay Kit below.

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