I'm going to perform drug treatment to my normal human epidermal keratinocyte cells (NHEKa). The drugs I'm using are anti-EGFRs and doxycycline.
Should I collect them right after the drug treatment or should i remove the old supernatant after the drug treatment and collect the ones reincubated with fresh medium? Why and why not to do so?
And how long can the supernatant be stored under -20 degree celcius condition?
Hey Phylia,
Although I am not an expert in cell culturing and performing ELISA's, from what I have been doing through my thesis project, I know that supernatant or the cell culture media needs to be collected after treatment (depending on your incubation time). This is done because after treatment cells are going to release substances like cytokines and other biomarkers into the cell media, so if you throw this away, it's like you threw away your effort.
So, after treatment collect your media in new ELISA plates and store them in the freezer until required. The remaining cells attached to the plate surface can be used for other procedures such as cell viability, qPCR, and others.
I hop this helps you and if I'm wrong, I am open to constructive criticism.
Thank you
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