A ProcartaPlex immunoassay (Thermo Fisher Scientific, Waltham, Massachusetts, USA) was used to assess the levels of 22 rat inflammatory markers (IL-1 alpha, G-CSF, IL-10, IL-17A, IL-1 beta, IL-6, TNF alpha, IL-4, GM-CSF, IFN gamma, IL-2, IL-5, IL-13, IL-12p70, eotaxin, GRO alpha, IP-10, MCP-1, MCP-3, MIP-1 alpha, MIP-2 and RANTES) in single muscle tissue samples. Sample preparation, assays and analyses were performed as described in the manufacturer’s instructions.
Procartaplex immunoassay
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ProcartaPlex Immunoassay is a multiplex assay platform that enables the simultaneous detection and quantification of multiple analytes in a single sample. It provides a powerful tool for researchers to measure various biomolecules, such as proteins, cytokines, and other targets, in a wide range of sample types.
Automatically generated - may contain errors
Lab products found in correlation
Procartaplex analyst 1, by Thermo Fisher Scientific (3 mentions)
Bio plex 200 instrument, by Bio-Rad (2 mentions)
Bio plex 200 system, by Bio-Rad (2 mentions)
Bead mill homogenizer, by Qiagen (1 mentions)
Mouse inflammation cytometric bead array cba, by BD (1 mentions)
Hydroxyproline assay kit, by Merck Group (1 mentions)
Luminex, by Thermo Fisher Scientific (1 mentions)
Facsaria 2, by BD (1 mentions)
Gm csf, by Sanofi (1 mentions)
Bio plex manager 6, by Bio-Rad (1 mentions)
Celltrace violet ctv, by Thermo Fisher Scientific (1 mentions)
Brefeldin a, by Merck Group (1 mentions)
Elisa, by Mabtech (1 mentions)
Poly 1 c, by InvivoGen (1 mentions)
Streptomycin strep, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Procartaplex universal assay buffer, by Thermo Fisher Scientific (1 mentions)
Flexmap 3d, by DiaSorin (1 mentions)
Tissuelyser 2, by Qiagen (1 mentions)
Bradford assay, by Bio-Rad (1 mentions)
48 protocols using procartaplex immunoassay
1
Comprehensive Analysis of Rat Inflammatory Markers
2
Influenza A/x31 Induced Lung Cytokines
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Lungs were harvested from Nkx2.1creTpl2flox/- mice, placed in 1 ml PBS and dissociated with a bead mill homogenizer (Qiagen). Lung homogenates were centrifuged at 500 x g for 5 min. Cytokines were quantified using a custom Procartaplex Immunoassay (Thermo-Scientific), IFN-α and IFN-β Luminescent ELISA kits (Invivogen) and Mouse Inflammation Cytometric Bead Array (CBA, BD Bioscience). IFN-λ3 (IL-28B) was measured using a colorimetric ELISA (Invitrogen). Hydroxyproline assay kit (Sigma, MAK357) was utilized to determine collagen levels in Nkx2.1creTpl2flox/- lungs infected with influenza A/x31 for 8 days prior to harvesting.
3
Cytokine and Growth Factor Analysis
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After 2 and 6 days of storage at 25 °C, immediately after the resazurin assay, the cell-seeded constructs were transferred to 1 mL of fresh CCM and were incubated at 37 °C/5% CO2. After 24 h, the conditioned medium was collected and was stored at −80 °C until use. The concentrations of the growth factors and cytokines (SDF-1α, VEGF-A, VCAM-1, FGF-2, HGF, MCP-1, IL-6, and bNGF) were determined using the Luminex®-based multiplex ProcartaPlex® Immunoassay (Thermo Fisher Scientific). Nine samples were measured for each time interval. CMs collected on day 8 from the same samples that were later measured after 2 and 6 days of storage at 25 °C served as controls. The measurement was performed on a Bio-Plex 200 Instrument (Bio-Rad, Prague, Czech Republic) according to the manufacturer’s instructions. All samples were analyzed in duplicate. The cytokine and growth factor concentrations (pg/mL or ng/mL) in the samples were derived from measured MFIs using fitted standard curves.
4
Quantifying IFNα and IFNβ in Serum
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IFNα and IFNβ levels in serum were measured using a magnetic bead-based ProcartaPlex Immunoassay (Thermo Fisher, Waltham, MA, USA) using manufacturers protocols.
5
Allogeneic MLR Cytokine Profiling
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For the allogeneic MLRs, CD1a+Langerin+ and CD1a+Langerin− cells were sorted on the basis of CD1a and Langerin expression with a FACS Aria II (BD Biosciences). Cells were cultured in the presence or absence of the cytokine maturation cocktail for 24 hours, both supplemented with 50 ng/ml GM-CSF (Sanofi), followed by coculturing with 0.4 μM carboxyfluorescein succinimidyl ester‒labeled (Thermo Fisher Scientific) allogeneic CD14− peripheral blood lymphocytes in the ratios of 1:5, 1:10, and 1:20 for 5 days. On day 5, supernatants from MLRs were collected, and cells were harvested to determine T cell proliferation by carboxyfluorescein succinimidyl ester dilution by flow cytometry. Supernatants were used for a ProcartaPlex Immunoassay (Thermo Fisher Scientific) for measurements of the cytokines IL-2, IL-4, IL-5, IL-9, IL-10, IL-13, IL-17A, and IFN-γ.
6
Multiplex Inflammatory Profiling in Rat Muscle
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A ProcartaPlex immunoassay (Thermo Fisher Scientific, Waltham, Massachusetts, USA) was used to assess the levels of 22 rat inflammatory markers (IL-1 alpha, G-CSF, IL-10, IL-17A, IL-1 beta, IL-6, TNF alpha, IL-4, GM-CSF, IFN gamma, IL-2, IL-5, IL-13, IL-12p70, Eotaxin, GRO alpha, IP-10, MCP-1, MCP-3, MIP-1 alpha, MIP-2, and Rantes) in single muscle tissue samples. Sample preparation, assays, and analyses were performed as described in the manufacturer's instructions.
7
Bispecific Antibodies and Cytokine Release
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Example 20
This example illustrates cytokine release in the presence of bispecific antibodies in vitro.
Human PBMCs from three donors were incubated with isotype, anti-CD3 antibody (OKT3, as positive control), and three bispecific antibodies (anti-PD-L1 #6, anti-Her2 #3-7, and anti-glycan conjugated to CD137 #54 scFv) at 0.67, 6.67, and 66.67 nM for 24 hours. Cytokines released into the culture medium were detected by the multiplex ProcartaPlex Immunoassay (ThermoFisher Scientific). A profound cytokine release was induced by OKT3, while the three bispecific antibodies did not induce cytokine release (
These results show that, compared to OKT3, PD-L1 #6-CD137 #54, Her2 #3-7-CD137 #54 and glycan-CD137 #54 bsAbs did not induce noticeable cytokine release above background when incubated with human PBMC.
8
Cytokine and Chemokine Profiling in Irradiated BAL
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Cytokine and chemokine levels in gamma irradiated (2 megarads) BAL specimens were analyzed using a custom ProcartaPlex immunoassay (ThermoFisher Scientific) nonhuman primate cytokine magnetic bead panel premixed 24-plex kit, per the manufacturer’s protocol. The levels of the analytes were assessed on a Bio-Plex 200 system (Bio-Rad).
9
Quantifying IFN-β Serum Levels
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IFN-β serum levels were assessed basally and after treatment using a customized Procartaplex Immunoassay (Thermo Fisher). 25 µL per well of antibody coupled beads and 50 µL per well of diluted patient serum were added into a 96 well plate and incubated overnight at 4°C. Each assay included a standard curve and all samples were performed in duplicate. Plates were processed following manufacturers’ instructions and then read with a Bio-Plex® 200 system (Bio-Rad). The data obtained were processed with the Bioplex Manager 6.0 software (Bio-Rad) using a five-parameter curve fitting algorithm to analyse them.
10
Quantifying Systemic Inflammatory Markers
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To analyze systemic inflammatory mediators, plasma from mice was analyzed by using the ProcartaPlex Immunoassay (ThermoFisher, Waltham, MA, USA) for granulocyte-colony stimulating factor (G-CSF), interleukin (IL)-6, keratinocyte chemoattractant (KC), IL-10, tumor necrosis factor (TNF), CXCL10 and monocyte chemoattractant protein-1 (MCP-1). All procedures were performed according to the manufacturer’s instructions. Some plasma parameters have been published as control samples in a previous study regarding cardiac inflammation after trauma (43 (link)).
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