The topic of my project is an immunosensor. For that, I dropped casted antibody with nanomaterial on the working electrode, then I kept the electrode at 4 c, but I had to keep that electrode at room temperature when I needed to take readings of CV, DPV, and EIS. So my question is, are antibodies stable at room temperature, then? What am I supposed to do if not? a similar question is also related to antigens. how long antibody and antigens stable at room temperature on the working electrode?
Yes, antibodies and most antigens are quite stable at room temperature. After all, they are able to function inside your body. Stability problems are usually due to microbial contamination. Thus, while scientists commonly perform immunochemical assays at room temperature for a few hours, for overnight incubations you should use 4C or include an antimicrobial. Sodium azide is commonly used, inexpensive, rapidly effective, and highly toxic to practically everything. In my experience 0.02% NaN3 is sufficient to maintain sterility. However, it destroys horseradish peroxidase, so it is incompatible with immunoassays utilizing this reporter enzyme. For HRP assays substitute 0.02% thimerosal.
John Carter
Sir,
To detect Anti "CA 15-3," I'm using "CA 15-3." During CV and EIS analysis, are they stable and alive at room temperature?
I'm not familiar with those particular proteins. Room temperature does not typically denature proteins. But if you observe a rapid falloff in signal as your assay warms to RT, you may have an atypically temperature sensitive Ag.
OTOH, if you observe generally poor sensitivity, then your proteins may be losing structure/activity upon immobilization. This frequently is a problem for Ag, and sometimes also for Ab. It's good to use an Ab-Ag pair that is validated for your assay type. What works well on a Western blot may not work at all on ELISA. And it's best to use an immobilization strategy that orients the proteins for optimum binding.
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