The levels of immunoreactive MCP-1 in the hRPE conditioned media, collected after 24 hr incubations of hRPE cells seeded into 24-well plates, were determined by modification of a double ligand ELISA method as previously described (Bian et al., 2004 (link)). Briefly, 200ul of non-concentrated conditioned medium from each culture well were distributed in 50ul aliquots in 4 wells of a 96-well ELISA plate and diluted 1:1 with buffer as per manufacturer eBioscience (San Diego, CA; Ray Biotech Norcross, GA; Pierce Biotechnology, Inc., Rockford, IL) instructions. For IL-1β, a commercial human IL-1β high sensitivity ELISA kit from eBioscience was used to detect IL-1β from 0 to 10pg/ml in conditioned media. For IL-18, human IL-18 ELISA kit from Ray Biotech was used to detect IL-18 from 0.5 to 75 pg/ml in the conditioned media. For low concentrations of MCP-1, the Endogen Human MCP-1 ELISA kit from Pierce Biotechnology, Inc. was used. In all assays the samples used were conditioned media without any pooling or concentrating. Standards included half-log dilutions of corresponding chemokines at concentrations from 1 pg to 100ng/well. Production of each cytokine per cell/per hr can be calculated by normalizing the reported cytokine values by dividing by 1 × 105 (cell seeding density per well)/24hr.
Human il 1β high sensitivity elisa kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Human IL-1β high sensitivity ELISA kit is a laboratory tool used to quantitatively measure the levels of interleukin-1 beta (IL-1β) in human samples, such as serum, plasma, cell culture supernatants, and others. This kit uses the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify IL-1β with high sensitivity.
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Lab products found in correlation
2 protocols using human il 1β high sensitivity elisa kit
1
Quantification of Inflammatory Cytokines in hRPE Conditioned Media
2
Inflammatory Molecule Serum Analysis
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To analyse serum levels of endotoxin and IL-1β, which are inflammatory molecules, the blood aliquoted in the SST on the day of the visit was centrifuged at 3,500 rpm and 22°C for 10 minutes, then placed in a 2-mL tube and stored at −80°C until a quantitative analysis was conducted. To prevent deformation of samples, the serum samples were dissolved on ice after which an enzyme-linked immunosorbent assay (human IL-1β high-sensitivity ELISA kit; eBioscience, San Diego, CA USA) was conducted and a quantitative analysis using the Pyrotell®-T kit was performed using the turbidimetric method (Pyros Kinetix®; Associates of Cape Cod Inc., East Falmouth, MA, USA).
All microbiological and immunological laboratory procedures were performed by blinded analysts, who did not have any knowledge of the clinical status of the study subjects.
All microbiological and immunological laboratory procedures were performed by blinded analysts, who did not have any knowledge of the clinical status of the study subjects.
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