The relative concentrations of cytokines were quantified (pg/ml) in macrophage culture supernatants after 48 h of stimulation using commercially available ELISA kits. Besides, wound beds and dorsal skin surrounding the wound were incised and cut into pieces in 600 μL PBS solution. The pieces and the solution were transferred in centrifuge tubes and centrifuged at 300 g for 5 min. The supernatants were collected and kept at −80 °C until further analysis. ELISAs for rat tumour necrosis factor-α (TNF-α), interleukin (IL)-10 (ELISA Development System, R&D Systems, Minneapolis, USA) and rat transforming growth factor-β1 (TGF-β1) (ELISA Development System, R&D Systems, USA) were performed according to the manufacturer's instructions.
Elisa development system
Manufactured by R&D Systems
Sourced in United States, United Kingdom
The ELISA (Enzyme-Linked Immunosorbent Assay) Development System is a laboratory instrument designed to perform sensitive and quantitative immunoassays. It provides the necessary components and protocols to develop and optimize ELISA-based assays for the detection and measurement of various analytes, such as proteins, peptides, hormones, and other biomolecules.
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Lab products found in correlation
6 protocols using elisa development system
1
Cytokine Quantification in Macrophage Cultures
2
Microglia Response to Cerebrospinal Fluid
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The BV2 cell line was derived from immortalised murine neonatal microglia and grown in 10% foetal bovine serum and 1% L-Glutamine supplemented Dulbecco’s Modified Eagle’s (DMEM) medium. The cells were incubated at 37 °C in a humidified atmosphere of 5% CO2 and 95% air, until the cell density reached approximately 1.6 × 106 cell/mL. 200 μL of CSF was diluted in 1 mL of DMEM and added to BV2 microglia cells. Every 24 h the supernatant was removed for analysis and replaced by fresh diluted CSF. The TNF-α concentration in the supernatant was quantified using the Duoset® enzyme-linked immunosorbent assay (ELISA) development system (R&D Systems, Abingdon, Oxfordshire, UK). Three wells for each CSF were used to estimate variation in the experiments. For antibody experiment, CSF and antibody were diluted in DMEM buffer and added to the vesicle containing glass coverslip.
3
Quantification of Inflammatory Markers
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The concentrations of MPO, MMPs and the cytokines IL-1β, IL-8, and tumor necrosis factor (TNF)-α in cell culture supernatants were measured using the DuoSet enzyme-linked immunosorbent assay (ELISA) development system (R&D Systems) and ELISA MAX Standard Set Human TNF-α (Cat# 430201; BioLegend, San Diego, CA, USA) according to manufacturer' instructions. The absorbance at 450 nm was measured on a microplate reader (2030 ARVO X4; PerkinElmer Japan; Tokyo).
4
Quantifying Inflammatory Cytokine Levels
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To determine cumulative TNF-α and Il-1β production, supernatants were obtained after incubation with the Aβ over viable time frames and stored at −80 °C until analysed. TNF-α, and Il-1β were analysed using the Duoset® enzyme-linked immunosorbent assay (ELISA) development system (R&D Systems, Abingdon, Oxfordshire, UK).
5
Serum Cytokine Profiles in Tuberculosis
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Serum CCL2 and IL-12p70, IFN-γ, TNF-γ, and TGF-β cytokines were assayed using respective human Duoset enzyme-linked immunosorbent assay (ELISA) Development System (R&D Systems, Minneapolis, MN, USA) in 120 tuberculosis cases (40 representative cases each from wild, heterozygous, and mutant genotypes) and 54 healthy controls (20 representative healthy controls each from heterozygous and wild genotypes and 14 from mutant genotypes with respect to the CCL2-2518A>G polymorphism).
6
Quantifying Inflammatory Markers in α-Syn Exposure
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To determine cumulative TNF-α, Il-1β and nitric oxide production, supernatants were obtained after incubation with the α-syn over viable time frames and stored at − 80° until analyzed. TNF-α and Il-1β were analyzed using the Duoset® enzyme-linked immunosorbent assay (ELISA) development system (R&D Systems, Abingdon, Oxfordshire, UK). iNOS activity was determined indirectly by measuring using the Griess reaction [18 (link)].
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