Hi, I am evaluating the toxicity of a mixture of agrochemicals and the possible induction of apoptosis. I used different concetrations and analyzed by the method of annexin V and I saw no significant differences in apoptotic cells. So I 'm looking for a positive control to see if this result depends on the technique , cells or treatment .
Which is a good positive control for apoptosis of A549 cells?
Thanks!
Hi Isis
My colleagues use staurosporine routinely for this and A549 is their preferred reference cell type. It would be useful to include a viability dye (eg DRAQ7) alongside Annexin V to help in the understanding of the cell death pathway being undergone by the cells. I have also seen the combination of FITC-Annexin V, TMRM (mitochondrial membrane potential) and DRAQ7 to give deeper temporal information. This combination can be used on flow cytometry or microscopy interchangeably and does not need UV laser excitation. It may further be possible to combine these in a real-time manner (see Akagi, J., et al., Cytometry Part A, Feb. 2013 - DRAQ7+TMRM - and Liang, J.R., et al. EMBO Reports 2013 - DRAQ7 + Annexin V) to better understand the impact of your cell treatment at more physiological dose / exposure.
Kind regards,
Roy
Maryam Majidinia Hi, may I know what percentage/concentration of DMSO did u use?
Munazza Fatima
Aapa, 24 hrs is the ideal time of incubation but you can also check fate of damage after 12 and 24 hrs of incubation as well !
You can try DNA damaging agents like Doxorubucin, Camptothecin, or Etoposide as positive controls.
actinomycin, puromycin, antimycin A, tetracycline, oligomycin, tamoxifen, dexamethasone
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