Assessment of IL-6 release was performed using CytoSet sandwich ELISA according to the manufacturer’s instructions (Life Technologies; Mulgrave, Vic, Australia). The limit of detection of the IL-6 assay was 16 pg/ml. The concentration of IL-1α and IL-1β in the media was performed by sandwich ELISA according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN). The limit of detection of the IL-1α and IL-1β assays was 7.8 and 2 pg/ml, respectively. All data were corrected for total protein and expressed as either pg or ng per mg protein. The protein content of tissue homogenates was determined using BCA protein assay (Pierce, Rockford, IL), using BSA as a reference standard, as previously described [21 (link)]. The calculated inter-assay and intra-assay coefficients of variation (CV) were all less than 10%.
Cytoset sandwich elisa
Manufactured by Thermo Fisher Scientific
Sourced in Australia
CytoSet sandwich ELISA is a laboratory equipment product designed for the detection and quantification of target analytes in biological samples using an enzyme-linked immunosorbent assay (ELISA) format. The product enables the capture and detection of target proteins in a sandwich-style ELISA configuration.
Automatically generated - may contain errors
4 protocols using cytoset sandwich elisa
1
Quantification of Cytokine Levels
2
Cytokine Release Quantification
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Assessment of IL-6 and IL-8 cytokine release was performed using CytoSet™ sandwich ELISA according to the manufacturer's instructions (Life Technologies, Carlsbad, CA). The limit of detection of the IL-6 and IL-8 assays was 16 and 12 pg/ml, respectively. The release of MCP-1 and GRO-α was performed using DuoSet™ sandwich ELISA according to the manufacturer's instructions (R&D Systems Inc., Minneapolis, MN). The limit of detection of GRO-α and MCP-1 assay was 31 and 15 pg/ml, respectively. The inter- and intra-assay coefficients of variation (CV) for all assays were less than 10%.
3
Cytokine and Angiogenic Factor Quantification
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IL6 and CXCL8 cytokine release was assessed using CytoSet sandwich ELISA according to the manufacturer's instructions (Life Technologies). The release of sFlt, CXCL1, CCL2 and IL1B was performed using DuoSet sandwich ELISA according to the manufacturer's instructions (R&D Systems). The inter-and intra-assay coefficients of variation for all assays were less than 10%.
4
Cytokine and Chemokine Release Assays
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Assessment of cytokine and chemokine release of IL6 and CXCL8 was performed using the CytoSet sandwich ELISA according to the manufacturer's instructions (Life Technologies). The release of CCL2, 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 (Cayman Chemical Company). The interassay and intraassay coefficients of variation for all assays were less than 10%.
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