Pelikine compact

Manufactured by R&D Systems

The PeliKine Compact is a laboratory equipment product manufactured by R&D Systems. It is designed to perform specific functions in a research setting, but a detailed and unbiased description of its core function cannot be provided without the risk of interpretation or extrapolation.

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Lab products found in correlation

Duoset elisa, by R&D Systems (2 mentions) Round bottom 96 well plate, by Greiner (2 mentions) Vacutainer tube, by BD (1 mentions) Pan monocyte isolation kit, by Miltenyi Biotec (1 mentions) Ficoll paque, by Avantor (1 mentions) Triton x 100, by Merck Group (1 mentions) Xn 450 hematology analyzer, by Sysmex (1 mentions)

7 protocols using pelikine compact

Cytokine Measurement in Cell Stimulation

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Samples of all experiments were measured at once using the following ELISA kits: In the 24-hour PBMC stimulation experiments we measured concentrations of human IL-1β, IL-6 as well as TNFα (PeliKine Compact or R&D Systems). Supernatants of the 7 days stimulation assays were used to measure IL-22, IL-17 or IFN-γ (PeliKine Compact or R&D Systems). For the whole blood samples IL-6, TNFα, IL-1β as well as IFN-γ levels were determined (PeliKine Compact or R&D Systems). Supernatants from the macrophage stimulation experiment were used for IL-6 and TNFα measurements (PeliKine Compact or R&D Systems).

ter Horst R., Jaeger M., Smeekens S.P., Oosting M., Swertz M.A., Li Y., Kumar V., Diavatopoulos D.A., Jansen A.F., Lemmers H., Toenhake-Dijkstra H., van Herwaarden A.E., Janssen M., van der Molen R.G., Joosten I., Sweep F.C., Smit J.W., Netea-Maier R.T., Koenders M.M., Xavier R.J., van der Meer J.W., Dinarello C.A., Pavelka N., Wijmenga C., Notebaart R.A., Joosten L.A, & Netea M.G. (2016). Host and environmental factors influencing individual human cytokine responses. Cell, 167(4), 1111-1124.e13.

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Measuring Cytokine Levels in PBMC Stimulation

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In the supernatants of the 24‐h PBMC stimulation experiments, concentrations of IL‐1β, IL‐6 and TNF‐α were measured following the manufacturer's protocols (PeliKine Compact or R&D Systems). Supernatants of the 7‐day stimulation assays were used to measure IL‐22, IL‐17 or IFN‐γ (PeliKine Compact or R&D Systems).

Jaeger M., Sloot Y.J., Horst R.T., Chu X., Koenen H.J., Koeken V.A., Moorlag S.J., de Bree C.J., Mourits V.P., Lemmers H., Dijkstra H., Medici M., van Herwaarden A.E., Joosten I., Joosten L.A., Li Y., Smit J.W., Netea M.G, & Netea‐Maier R.T. (2021). Thyrotrophin and thyroxine support immune homeostasis in humans. Immunology, 163(2), 155-168.

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Cytokine Profiling of Immune Cells

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Venous blood was collected in sterile 10 mL EDTA and 8 mL serum BD Vacutainer tubes (Becton Dickinson) and processed within 1–4 hours. Isolation of PBMCs was performed on freshly collected blood by density centrifugation over Ficoll-Paque (VWR) as described previously (70 (link)). Monocytes were isolated by magnetic-activated cell sorting using negative bead selection with the Pan Monocyte Isolation Kit (Miltenyi Biotec) according to manufacturer’s instructions. Cell counts, cell purity, and composition were evaluated by XN-450 hematology analyzer (Sysmex Corporation). Fresh isolated cells (monocytes 1 × 10⁵ cells per well, PBMCs 5 × 10⁵ cells per well) were incubated with different bacterial, fungal, and viral stimuli (Supplemental Table 2) at 37°C and 5% CO₂ for either 24 hours or 7 days. For the 7-day stimulation, 10% human pooled serum was added to the wells. IL-1β, IL-6, IL-1Ra, IL-10, and TNF-α were determined in the supernatants of the 24-hour PBMC or monocyte stimulation experiments, using ELISAs (Duoset ELISA, R&D Systems). IL-17, IL-22, and IFN-γ were measured after the 7-day stimulation of PBMCs (PeliKine Compact or R&D Systems). For intracellular cytokine measurements of active IL-1β and pro–IL-1β (Quantikine, R&D Systems), cell pellets were lysed using Triton X-100 (MilliporeSigma).

van der Heijden W.A., Van de Wijer L., Keramati F., Trypsteen W., Rutsaert S., ter Horst R., Jaeger M., Koenen H.J., Stunnenberg H.G., Joosten I., Verweij P.E., van Lunzen J., Dinarello C.A., Joosten L.A., Vandekerckhove L., Netea M.G., van der Ven A.J, & de Mast Q. (2021). Chronic HIV infection induces transcriptional and functional reprogramming of innate immune cells. JCI Insight, 6(7), e145928.

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Cytokine profiling of ex vivo pathogen stimulation

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Heat-killed human pathogens and microbial ligands were used for ex vivo stimulation. Isolation of PBMCs from fresh blood was performed with gradient centrifugation as previously described. Cells were resuspended in RPMI-1640 complete culture medium. PBMC stimulations were performed with 5 × 10⁵ cells/well in round-bottom 96-well plates (Greiner) for either 24 h or 7 days in the presence of 10% human pooled serum (HPS) at 37 °C and 5% CO₂. Whole blood was stimulated for 48 h. Supernatants were collected and stored at −20 °C until cytokine measurement. Concentrations of Interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, IL-17, IL-22 and interferon (IFN)-γ were measured following the manufacturer's protocols (PeliKine Compact or R&D Systems). The full list of cytokines, stimuli, cell systems and incubation times is provided in Supplementary data (Supplementary Table S2).

Peng C., Jiang X., Jaeger M., van Houten P., van Herwaarden A.E., Koeken V.A., Moorlag S.J., Mourits V.P., Lemmers H., Dijkstra H., Koenen H.J., Joosten I., van Cranenbroek B., Li Y., Joosten L.A., Netea M.G., Netea-Maier R.T, & Xu C.J. (2023). 11-deoxycortisol positively correlates with T cell immune traits in physiological conditions. eBioMedicine, 99, 104935.

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Serum IL-6 Levels in Candidemia Patients

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Concentrations of human IL-6 in the serum of candidemia patients were determined using specific commercial ELISA kits (PeliKine Compact or R&D Systems), in accordance with the manufacturers’ instructions. The data were available for 117 Caucasian candidemia patients. Candidemia patients were stratified on rs7036187 SNP genotype to obtain 111 AA and 6 AG patients. For each individual the median values of IL-6 levels measured across 15 days were used. The statistical difference was tested using a student t test (one-sided) by comparing the log2 transformed IL-6 values. P value less than 0.05 was considered significant.

Li Y., Oosting M., Deelen P., Ricaño-Ponce I., Smeekens S., Jaeger M., Matzaraki V., Swertz M.A., Xavier R.J., Franke L., Wijmenga C., Joosten L.A., Kumar V, & Netea M.G. (2016). Inter-individual variability and genetic influences on cytokine responses against bacterial and fungal pathogens. Nature medicine, 22(8), 952-960.

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Cytokine QTL Mapping Protocol

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Following the generation of genotype and cytokine data, cytokine QTLs were mapped as described previously (43 (link)). Briefly, concentrations of human cytokines were determined using specific commercial enzyme-linked immunosorbent assay (ELISA) kits (PeliKine Compact, or R&D Systems), per the manufacturer’s instructions. Cytokine QTLs were identified by log-transforming raw cytokine levels and mapping them to genotype data using a linear regression model with age, gender, and cell counts as covariates.

Gonçalves S.M., Antunes D., Leite L., Mercier T., ter Horst R., Vieira J., Espada E., Pinho Vaz C., Branca R., Campilho F., Freitas F., Ligeiro D., Marques A., van de Veerdonk F.L., Joosten L.A., Lagrou K., Maertens J., Netea M.G., Lacerda J.F., Campos A J.r., Cunha C, & Carvalho A. (2021). Genetic Variation in PFKFB3 Impairs Antifungal Immunometabolic Responses and Predisposes to Invasive Pulmonary Aspergillosis. mBio, 12(3), e00369-21.

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Cytokine Production in Immune Cell Assay

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Freshly isolated PBMCs were incubated with different stimuli (Supplementary Table 2) including bacterial (Staphylococcus aureus, M. tuberculosis, Streptococcus pneumoniae [S. pneumoniae]), fungal (Cryptococcus gattii, Candida albicans [C. albicans] hyphae and yeast) and other relevant antigens (Imiquimod, TLR7 ligand), in round-bottom 96-well plates (Greiner Bio-One, Frickenhausen, Germany) with 0.5 x 10⁶ cells/well at 37°C and 5% CO₂ in the presence of 10% human pooled serum for seven days. Supernatants were stored at -20°C. Levels of the lymphocyte-derived cytokines IL-17, IL-22, and interferon (IFN)-γ were measured in the supernatants (PeliKine Compact or Duoset ELISA, R&D Systems).

Van de Wijer L., van der Heijden W.A., ter Horst R., Jaeger M., Trypsteen W., Rutsaert S., van Cranenbroek B., van Rijssen E., Joosten I., Joosten L., Vandekerckhove L., Schoofs T., van Lunzen J., Netea M.G., Koenen H.J., van der Ven A.J, & de Mast Q. (2021). The Architecture of Circulating Immune Cells Is Dysregulated in People Living With HIV on Long Term Antiretroviral Treatment and Relates With Markers of the HIV-1 Reservoir, Cytomegalovirus, and Microbial Translocation. Frontiers in Immunology, 12, 661990.

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