I am currently testing multiplex systems (luminex, spx, and others) for cytokine assays. We would like to spike human serum with known concentrations of various cytokines and use those samples across all platforms to determine accuracy and precision. NIBSC is one source of the cytokines but I wanted to know if there are others that are recommended (bd, fisher, r and d) Does anyone have any experience in this area and can offer some tips and tricks? Thanks!
I can recommend Immunotools. They have a wide range of cytokines, are affordable and I never had any problems.
We use bioplex from Bio rad. It works realy well and I have done upto 10 plexes, however it can do as many as 500 analytes in a single sample. You do need a dedicated instrument for this though.
Biolegend also offers custom panels of several cytokines, which you can analyze using a flow cytometer.
Best
An update to the investigation I was working on... I ordered recombinant cytokines from NIBSC. They have approximate mass listed for each, and concentration is in IU. We report in pg/ml, so I dissolved the 1ug of lyophilized powder of each cytokine in 1ml pbs/01.%bsa. I then spiked serum, synthetic serum, and pbs/bsa in concentrations of 10pg/ml up to 4000 pg/ml. This corresponded to 1IU to 400IU. I then plotted pg/ml readouts from each instrument against intended IU concentrations, obtained a linear equation from that plot, and back solved for IU recovered. This helped us to determine linearity and spike recovery of each assay type for each cytokine tested. Hope this helps someone else in trying to determine w/v concentration from IU in the future!
Hi Kristen Ciano, thank you for your question.
I have added a link and attachment for potential vendor sources.
https://www.luminexcorp.com/blog/need-antibodies-for-that-luminex-assay-try-these-resources-to-find-potential-suppliers-and-user-reviews/
Hope this paper helps you:
Article Effects of serum and plasma matrices on multiplex immunoassays
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