Human IL-6 Uncoated ELISA Kit - FAQs

A: It is critically important to use a high-affinity binding PVDF membrane plate, rather than regular ELISA plate or nitrocellulose membrane plate. Additionally, highest affinity antibodies for capture work best in this assay. Empirical optimization of conditions such as cell density, stimulus/mitogen, and kinetics is necessary.

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We do not recommend the use of our ELISA kits for the ELISPOT application. We cannot guarantee the successful use of our ELISA kits in an ELISPOT application.

Find additional tips, troubleshooting help, and resources within our Antibodies and Immunoassays Support Center.

Most, but not all, of our ELISA antibody pairs work for ELISPOT. We continue to evaluate and optimize our anti-human and anti-mouse cytokine antibody pairs for ELISPOT applications. Please see our protocols page (www.thermofisher.com/us/en/home/references/protocols/cell-and-tissue-analysis/protocols) for a detailed protocol and a reference table with the list of antibody pairs that can be used in ELISPOT.

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Capture Antibody: Pre-titrated, Functional Grade (low endotoxin) purified antibody
Detection Antibody: Pre-titrated, biotin-conjugated antibody
10X Sterile Azide-Free Coating Buffer
5X ELISA/ELISPOT Diluent
Detection enzyme: pre-titrated Avidin HRP
Certificate of Analysis: Lot-specific instructions for dilution of antibodies and enzyme

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.