horizontal gel electrophoresis platform and covered with pH > 12 alkaline solution for 20 min

700 WordsApr 23, 20193 Pages
horizontal gel electrophoresis platform and covered with pH > 12 alkaline solution for 20 min without electrophoresis[15]. The slide were then neutralized and stained with 50 µl of 20 µg /ml ethidium promide. This cytotoxicity method of cell growth following to pro-mutagen exposure was detected under cytocharasin B and frequency binuclear cells. Fairbrain and co-workers reported the advantage of this technique include: i) high sensitivity for detecting low levels of DNA damage; ii) ability to detect genotoxicity in the absence of mitotic activity; iii) the requirement for small numbers of cells per sample; iv) flexibility; v) low costs; vi) easy application and vii) the relatively short time period needed to complete an experiment [16].…show more content…
The plate was then read for absorbance at 540nm using the microplate reader. The neutral red assay was chosen because it assesses the lysosomal integrity of cells. As the integrity of the lysosome in cells is dependent upon a very wide range of healthy internal machinery, as opposed to a single or a few individual pathways, it provides a good overview of general cellular health as deficient systems inside the cell are likely to impact on the cells ability to retain the neutralred stain [19] . LDH release Lactate dehydrogenase (LDH) is an oxidative enzyme that changes lactate into pyruvate during glycolysis. LDH widely exists in cell membranes and cytoplasm, and is released from cells into culture supernatants immediately after cell damage. Therefore, photospectrometrical assessment for cell viability through the extracellular leakage of LDH can be applied for the antiviral evaluation of compounds [20] . The method using cell line and described previously. The LDH released from necrotic cells into the extracellular fluid was determined after 4 h of treatment with the test agents by using the commercially available Cytotoxicity Detection Kit (LDH). The plates were centrifuged at 400 g and 4 ◦C for 4 min and an aliquot of 50 μl was taken to quantify the LDH[21]. Adenosine triphosphate (ATP) content Adenosine triphosphate (ATP), that is present in all metabolically active cells, can be determined in a bioluminescent
Open Document