IL-1β in microglia culture supernatant was measured with Mouse IL-1β /IL-1F2 DuoSet ELISA Kit (R&D Systems DY401, Minneapolis MN) according to the manufacturer's instructions. CXCL10 was measured in neuronal culture supernatant with Mouse CXCL10 DuoSet ELISA (R&D Systems DY466-05, Minneapolis MN) according to the manufacturer's instructions. Primary microglia were stimulated with recombinant Mouse CXCL10 protein (R&D Systems 466-CR-608 050/CF, Minneapolis MN) and incubated with Mouse CXCL10 antibody (R&D Systems AF-466-609 NA, Minneapolis MN). TNF-α and IL-1β were measured in the supernatant by ELISA with Mouse TNF-α Quantikine ELISA Kit (R&D Systems MTA00B, Minneapolis MN) and Mouse IL-1β/IL-1F2 DuoSet ELISA Kit according to manufacturer's instructions.
Mouse il 1β il 1f2 duoset elisa kit
Manufactured by R&D Systems
Sourced in United States
The Mouse IL-1β/IL-1F2 DuoSet ELISA kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of mouse interleukin-1 beta (IL-1β) and interleukin-1 family member 2 (IL-1F2) levels in cell culture supernatants and serum samples.
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Lab products found in correlation
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Precellys evolution instrument, by Bertin Technologies (1 mentions)
Mouse tnf α duoset elisa kit, by R&D Systems (1 mentions)
Cytotox 96 non radioactive cytotoxicity assay, by Promega (1 mentions)
4 protocols using mouse il 1β il 1f2 duoset elisa kit
1
Cytokine Measurements in Neuroinflammation
2
Measuring Mouse Interleukin-1β Levels
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Interleukin-1β (IL-1β) protein level was measured by mouse IL-1β/IL-1F2 DuoSet ELISA Kit from R&D Systems (DY401, Minneapolis, MN, United States) according to the manufacturers’ instructions.
3
Quantification of Inflammatory Cytokines in Allergen-Challenged Skin
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IL-1β, IL-6 and TNFα were quantified from full ear skin samples (dermis + epidermis) following allergen re-exposure. The skin was removed from the cartilage tissue using surgical tweezers, and snap-frozen in liquid nitrogen 24 h post challenge. Samples were submerged in 800 µL lysis buffer (50 mM Trizma base, 250 mM NaCl, 6.4 mM EDTA, 17.6 mM Triton X-100, pH 7.4, including protease inhibitor cocktail (cOmplete, Roche Diagnostics GmbH, Berlin, Germany)). Tissue homogenization was performed in MK28 hard grinding precellys tubes (Bertin Technologies, Montigny-le-bretonneux, France) using a Precellys Evolution instrument (Bertin Technologies). Protein concentration in the supernatant was quantified by a Bradford assay and stored at −80 °C. The protein concentration of all samples was adjusted to 2 mg/mL. IL-1β levels were analyzed using a mouse IL-1β/IL-1F2 DuoSet ELISA kit (R&D systems), on 10× diluted samples. IL-6 and TNFα levels were analyzed using respective ELISA MAX™ standard kits (Biolegend) on undiluted samples. All assays were performed following manufacturer’s protocol.
4
Quantifying Cytokine and Cytotoxicity Levels
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Cytokine levels in cell supernatants were assessed using the mouse IL-1β/IL-1F2 DuoSet ELISA kit (R&D Systems) and mouse TNF-α DuoSet ELISA kit (R&D Systems) according to the manufacturer’s instructions. ELISA plates were measured at OD450 using a microplate reader, and cytokine levels were quantified by interpolation using a standard curve. Supernatants were assayed for LDH release immediately after stimulation time courses per the manufacturer’s protocol from the CytoTox 96 Non-Radioactive Cytotoxicity Assay (Promega). Measurements of absorbance readings were performed on a microplate reader at wavelengths of 490 nm and 680 nm. LDH release was assessed as a percentage of background-subtracted maximum LDH values from lysed cells.
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