Thu Feb 20, 2014 4:16 pm by mleipold
Hi Vincent,
At the Stanford HIMC, we generally elute the protein in 100uL W buffer, spec it for protein concentration, then dilute it with an equal volume of antibody stabilizer (final 200uL total, at 50% of original concentration; 50/50 W buffer/stabilizer). Most of the time, we don't readjust the concentration after that, we just add more or less volume to the antibody cocktail if we have to re-titer.
At 50/50, we get reproducibly at least 4 months out of all of our antibody stocks. There are a few that start to decrease around 4-5 months, but we run enough experiments that we usually use the stock up before then. Many antibodies are stable for 6+ months.
According to the stabilizer buffer manufacturer, the more stabilizer, the better. They do recommend re-spinning after protein determination, and resuspending in 100% stabilizer. I know from talking with DVS, that this is what *they* do...all of their preconjugated antibodies are in 100% stabilizer. DVS has also done lifetime stabilities (accelerated, and regular), and they guarantee 6 months. However, I've heard that they're good for longer than that: DVS just doesn't want to officially guarantee that.
Mike