Cytokines in thawed plasma (dilution, 1:10) and lung lavage fluid (dilution, 1:2) were measured by using sandwich ELISA kits according to the manufacturer's instructions. Albumin was measured in thawed plasma (dilution, 1:250,000) and lung lavage fluid (dilution, 1:1000). The assays used and their lower limit of detection (LLD) were: Mouse IL-6 DuoSet ELISA kit (LLD, 15.6 pg/mL; R and D Systems, Minneapolis, MN); Mouse Intracellular Adhesion Molecule-1 (ICAM-1) DuoSet ELISA kit (LLD 125 pg/mL; R and D Systems); Mouse Receptor for Advanced Glycation End-products (RAGE) DuoSet ELISA kit (LLD, 125 pg/mL; R and D Systems); Mouse IL-1β Platinum ELISA (LLD, 7.8 pg/mL; Thermo Fisher); Mouse TNF-α Platinum ELISA (LLD, 31.3 pg/mL; Thermo Fisher); and Mouse Albumin ELISA kit (LLD, 1.23 ng/mL; Bethyl Laboratories, Montgomery, TX).Absorbance was read on a spectrophotometer (Biotek, Winooski, VT) at a measuring wavelength of 450 nm for all ELISAs except IL-1β and TNF-α (reference wavelength 630 nm).The data were analyzed by using KC4 software (Biotek).
Mouse il 1β platinum elisa
Manufactured by Thermo Fisher Scientific
Sourced in Ireland
The Mouse IL-1β Platinum ELISA is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of mouse interleukin-1 beta (IL-1β) levels in cell culture supernatants, serum, and plasma.
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2 protocols using mouse il 1β platinum elisa
1
Cytokine and Albumin Measurement in Biological Fluids
2
Measuring Macrophage Inflammatory Responses
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Supernatants were collected from BMDM following activation, and subsequently subjected to analysis using the Griess Assay system, according to manufacturer’s instructions (Promega, Southampton, UK, #G2930). This assay measures NO2-, which is a breakdown product of NO production and indicative of NO production by the macrophages.
ELISAs were performed using the Mouse IL-1β platinum ELISA and Mouse TNF-α Instant ELISA kits (#BMS6002, #BMS607/2INST, Thermofisher Scientific, Biosciences, Dublin, Ireland), following the manufacturer’s instructions. Signals were measured using a SpectroMax plate reader spectrophotometer coupled to SoftMaxPro Software (Molecular Devices, Wokingham, UK).
ELISAs were performed using the Mouse IL-1β platinum ELISA and Mouse TNF-α Instant ELISA kits (#BMS6002, #BMS607/2INST, Thermofisher Scientific, Biosciences, Dublin, Ireland), following the manufacturer’s instructions. Signals were measured using a SpectroMax plate reader spectrophotometer coupled to SoftMaxPro Software (Molecular Devices, Wokingham, UK).
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