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FAQs for V-PLEX Proinflammatory Panel 1 (mouse) Kits

  1. What is the difference between the 10-plex panel, a custom panel, and the single assay kits?

    All Proinflammatory Panel 1 (mouse) assays use the same capture and detection antibodies, diluents, and calibrators. All multiplex assays use a 10-spot plate; singleplex assays are provided on single spot plates. Each kit is provided with the specific detection antibodies for the assays ordered—10 individual detection antibodies come with the full panel, 2 to 9 with custom panels, and 1 with single assay kits.

  2. How do the assays in the Proinflammatory Panel 1 (mouse) compare to the assays in the previous mouse cytokine panel (Mouse ProInflammatory 7-Plex Ultra-Sensitive Kit)?

    • The assays in the new kit are more sensitive and have been tested in mouse cell culture supernatants, serum, plasma, urine, and stimulated cells in whole blood and in cell culture. The product insert contains data for all sample types. The new assays have also undergone multi-lot validation.
    • Where available, the calibrators are anchored to NIBSC or WHO reference material. If an NIBSC or WHO reference material is not available, the calibrator is anchored to an internal reference material. Calibrators are provided lyophilized in the new kit.
    • Controls are available and included in the V-PLEX Plus Kit.
    • The new kit is provided with a different set of diluents.

  3. Is cross-reactivity testing conducted for the Proinflammatory Panel 1 (mouse) Kit?

    Yes. Cross-reactivity is measured for each kit lot using individual detection antibodies. During development, we tested individual calibrators and individual detection antibodies to validate assay specificity. Non-specific binding was also evaluated with additional recombinant mouse analytes (IL-13, IL-17, GM-CSF, MCP-1, MIP-3α, RANTES, TNF-RI, TNF-RII, and VEGF). No significant crossreactivity was observed.

  4. For analytes that have binding partners such as corresponding receptors, would the assay measure free analyte only, bound analyte, or both?

    We tested the TNF-α assay for interference by its receptors, the Proinflammatory Panel 1 (mouse) was run with TNF-α–containing serum to which 15 ng/mL of TNF-RI or TNF-RII was added. Assay interference was not observed. Other soluble receptors have not been tested.

  5. Are the calibrators in the Proinflammatory Panel 1 (mouse) Kit from native protein?

    The calibrators are recombinant proteins expressed in E. coli or Sf21 cells.

  6.  Is it feasible to store the reconstituted highest calibrator for longer than 30 days? Would there be similar recommendations for reconstituted controls?

    Yes, the reconstituted highest calibrator or reconstituted controls can be stored frozen (≤ -70°C) when longer than 30 day storage is needed. Reconstituted calibrator and reconstituted controls can undergo 5 freeze-thaw cycles without significantly affecting the performance of the assay. Calibrator and controls that have been further diluted should be discarded.

  7. Can the assay protocol be further streamlined to accommodate automation?

    Yes, alternative protocols for reduced wash steps, extended incubation, and diluting samples in the plate are described in the product insert. Calibration curves comparing the recommended protocol to alternative protocols are shown in the product insert.

 





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