I want to determine cytotoxicity of oxidized LDL in THP-1 macrophages in culture (adherent cells after PMA treatment). Is the LDH release assay a good choice. Can I collect and freeze the supernatant from the THP-1 after ox LDL treatment and run the LDH release assay later? Is LDH release a marker of necrosis only and not a measure of apoptosis? For quantifying apoptosis plan to do caspase 3 Immunofluorescence.
Carbon nanodots interference with lactate dehydrogenase assay in human monocyte THP-1 cells
Petra C Wright, Hu Qin, Martin MF Choi, Norman HL Chiu, and Zhenquan Jia
Abstract
Background
Carbon nanodots (CD), a new class of carbon nanomaterials with sizes below 10 nm, have recently attracted wide attention due to their superiority in water solubility, chemical inertness, and resistance to photobleaching. As a result, CD has found important and wide applications in energy, catalysis, biological labeling, bioimaging and drug delivery. On the other hand, due to the lack of available toxicity data, there is a growing concern regarding the potential risks of CD. Hence, accurate assessment of the cytotoxicity of CD has become more important than ever before. The lactate dehydrogenase (LDH) assay is widely used to detect cytotoxicity of various nanoparticles including CD. Many recent studies used LDH assay to study the CD toxicity in various cells. However, these studies failed to further examine whether the CD were interfering with the LDH assay which would alter their findings.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4213343/
1. LDH is for cell membrane damage (always necrosis/late apoptosis), better for early necrotic compounds
2. Medium can be frozen
3. Proper (like 10Ex6/ml) cell number is essential
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