Hello everybody,
For a few weeks now, adding recombinant interferons (λ1, λ2, β, γ) to my cultured cells kills most of them in approximately 24h. I use peprotech produces, solubilized in a 0.1% BSA-milliQ water solution, at a 20 ng/ml concentration.
The thing is, until early March, I had no problem with the exact same cells and IFNs. Suddenly, it started going wrong... I have then changed my cells with fresh out-of-the-N ones, bought new IFNs, changed dishes, changed the guy holding the micropipet, changed incubator... It still wouldn't work !
Any idea people??? I really need some help here...
Dear friend,
I don't like to say that sometimes a few of our experiments don't hold on the Reliability condition in an experiment, I hope you don't engage with this one.
The thing that I can say is: please check concentrations twice, volume measurements being checked twice and "Care about 0.1% BSA-milliQ water solution ( are you sure is it Ok???)".
I hope you find the true pathway.
Amir
The human IFN-alphas we used to treat our primary cell cultures are at 100pg/ml level, 20ng/ml seems very high. And just sort let you know, every batch of IFNs are not necessarily the same in terms of concentration and potency. You might want to titer the potency before using a new batch. And aliquot it into smaller portion so that you don't freeze and thaw the whole thing, which can cause it to degrade and loose potency.
Kejun Guo I thought of that to, that's why I'm testing my IFNs at different concentrations today. However, I am used to these concentrations, without any problem... But as you say, it may vary widely in potency... I already alicot my IFNs in order to avoid repeat thawing... But indeed, I used old batches, and new ones might be far more potent an maybe toxic with my cells... I'll see !
- Badges
- Science topic
- Similar topics
- Biological Science
- Cell Biology
- Culture Cells