Assessment of cytokine and chemokine release of IL-6 and IL-8 was performed using the CytoSet™ sandwich ELISA according to the manufacturer's instructions (Life Technologies; Mulgrave, Vic, Australia). The release of MCP-1, sICAM-1, and sVCAM-1 was performed by sandwich ELISA from R&D Systems (Minneapolis, MN, USA) according to the manufacturer's instructions. The release of PGF2α into the incubation medium was assayed using a commercially available competitive enzyme immunoassay kit according to the manufacturer's specifications (Cayman Chemical Company; Ann Arbor, MI, USA). The interassay and intraassay coefficients of variation for all assays were less than 10%.
Cytoset
Manufactured by Thermo Fisher Scientific
Sourced in United States, Australia
CytoSet is a laboratory instrument designed for cell analysis and sorting. It is used to identify, count, and separate specific cell types within a sample.
Automatically generated - may contain errors
Lab products found in correlation
Sandwich elisa, by R&D Systems (3 mentions)
Kookaburra kits, by Sapphire Bioscience (3 mentions)
Nunc maxisorb plates, by Thermo Fisher Scientific (3 mentions)
Duoset, by R&D Systems (3 mentions)
Spectramax m2, by Molecular Devices (2 mentions)
Competitive enzyme immunoassay kit, by Cayman Chemical (1 mentions)
Novex, by Thermo Fisher Scientific (1 mentions)
Sunrise plate reader, by Tecan (1 mentions)
Simplestep elisa kit, by Abcam (1 mentions)
10 protocols using cytoset
1
Cytokine and Chemokine ELISA Assay
2
Cytokine and Adhesion Molecule Quantification
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IL-6 and IL-8 release were assessed using the CytoSet™ sandwich ELISA, as instructed (Life Technologies). MCP-1 and ICAM-1 release were assessed by sandwich ELISA from R&D Systems (Minneapolis, MN, USA), as instructed. The interassay and intra-assay coefficients of variation for all assays were less than 10%.
3
Cytokine and Prostaglandin Release Assay
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The release of IL6 and IL8 was performed using CytoSet sandwich ELISA according to the manufacturer's instructions (Life Technologies). The limit of detection of the IL6 and IL8 assays was 16 and 12 pg/ml respectively. The release of PGE 2 and PGF 2a into the incubation medium was assayed using a commercially available competitive enzyme immunoassay kit according to the manufacturer's specifications (Kookaburra Kits from Sapphire Bioscience, Waterloo, NSW, Australia). The limit of detection of the PGE 2 and PGF 2a assays was 16 and 60 pg/ml respectively. For all assays, the interassay and intraassay coefficients of variation were !10%.
4
Cytokine, Chemokine, and Adhesion Molecule Release Assays
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Assessment of cytokine and chemokine release of IL6 and CXCL8was performed using the CytoSet™ sandwich ELISA according to the manufacturer's instructions (Life Technologies;
Mulgrave, Vic, Australia). The release of sICAM1 and sVCAM1 was performed by sandwich ELISA from R&D Systems (Minneapolis, MN, USA) according to the manufacturer's instructions.
The release of PGF 2α into the incubation medium was assayed using a commercially available competitive enzyme immunoassay kit according to the manufacturer's specifications (Kookaburra Kits from Sapphire Bioscience, NSW, Australia). The interassay and intraassay coefficients of variation for all assays were less than 10%.
Mulgrave, Vic, Australia). The release of sICAM1 and sVCAM1 was performed by sandwich ELISA from R&D Systems (Minneapolis, MN, USA) according to the manufacturer's instructions.
The release of PGF 2α into the incubation medium was assayed using a commercially available competitive enzyme immunoassay kit according to the manufacturer's specifications (Kookaburra Kits from Sapphire Bioscience, NSW, Australia). The interassay and intraassay coefficients of variation for all assays were less than 10%.
5
Cytokine and Prostaglandin Release Assays
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The release of IL6, CXCL8 and chemokine (C-C motif) ligand 2 (CCL2) was performed using CytoSet sandwich ELISA, according to the manufacturer's instructions (Life Technologies). The limit of detection of the IL6, CXCL8 and CCL2 assays was 16, 12 and 15 pg/ml respectively. The release of PGF 2a into the incubation medium was assayed using a commercially available competitive enzyme immunoassay kit, according to the manufacturer's specifications (Kookaburra Kits from Sapphire Bioscience, Redfern, NSW, Australia). The limit of detection of the PGF 2a assay was 60 pg/ml. For all assays, the inter-and intra-assay coefficients of variation were !10%.
6
Quantifying CXCL8 and IL-1β in Cell Culture
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The concentrations of CXCL8 and IL-1β in the cell culture supernatant were measured using Cytoset (Invitrogen by Thermo Fischer Scientific, Waltham, MA, USA and Novex by Life Technologies Waltham, MA, USA) or Duoset (R&D systems, Minneapolis, MN, USA) enzyme-linked immunosorbent assay (ELISA) kits, respectively, both applied according to the manufacturer’s instructions. Samples were diluted in the recommended assay buffer to stay within the appropriate range of the assay. ELISA was performed in Nunc Maxisorb plates (Thermo Scientific, Waltham, MA, USA). Absorbance was measured using a Sunrise plate reader (Tecan, Männerdorf, Switzerland).
7
Cytokine Detection by ELISA
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Supernatants levels of TNF-α, IL-4, IL-10, IL-17 and IFN-γ were measured by ELISA (Cytoset, Invitrogen). Supernatants were diluted (1:5) before performing the assay. The absorbance was measured at 450 nm using the SpectraMax® M2 (Molecular Devices, Sunnyvale, CA).
8
ELISA Cytokine Quantification
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The concentrations of cytokines in the cell culture supernatants were measured using Cytoset (TNFα and CXCL8, Invitrogen by Thermo Fischer Scientific and Novex by Life Technologies) or Duoset (IL-1α and IL-1β, R&D systems, Minneapolis, MN, USA) enzyme-linked immunosorbent assay (ELISA) kits, both applied according to the manufacturer’s instructions. ELISA was performed in Nunc Maxisorb plates (Thermo Scientific, Waltham, MA, USA). Absorbance was measured using a Tecan Sunrise plate reader (Tecan, Männedorf, Switzerland).
9
Cytokine Profiling of Immune Cells
10
Cytokine and NLRP3 Expression Analysis
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The concentrations of cytokines in the cell culture supernatant were measured using Cytoset (TNFα and CXCL8; Invitrogen by Thermo Fischer Scientific, Waltham, MA, USA and Novex by Life Technologies, Waltham, MA, USA) or Duoset (IL-1α and IL-1β; R&D systems, Inc., Minneapolis, MN, USA) ELISA kits, while NLRP3 expression in cell lysates was determined using a SimpleStep ELISA kit (Abcam, Cambridge, United Kingdom). All kits were applied according to the manufacturer’s instructions. ELISA was performed in Nunc Maxisorb plates (Thermo Scientific, Waltham, MA, USA). Absorbance was measured using a Tecan Sunrise plate reader (Tecan, Männedorf, Switzerland).
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