To measure skin TSLP levels, ears were incubated overnight in culture media, and cell-free supernatants were stored. Measurement of TSLP was performed using a commercially available Duoset ELISA kit (eBioscience). Antigen-specific serum IgE responses were measured as described before. 30 (link) For histological analysis, tissues were fixed in 4% paraformaldehyde and embedded in paraffin, and sections were stained with hematoxylin and eosin (H&E). For real-time PCR analysis, RNA was isolated using an RNeasy mini kit (QIAGEN) according to the manufacturer’s instructions. cDNA was generated using a SuperscriptII reverse transcription kit (Invitrogen). Real-time quantitative PCR was performed on cDNA using SYBR green master mix (Applied Biosystems) and commercially available primer sets from Qiagen (Quantitect primer assays). Samples were normalized to β-actin and displayed as fold induction over controls.
Duoset elisa kit
Manufactured by Thermo Fisher Scientific
The Duoset ELISA kit is a complete set of reagents for the development of sandwich enzyme-linked immunosorbent assays (ELISA). It includes all the necessary components, such as capture and detection antibodies, standard, and substrate solution, to establish a sensitive and specific ELISA for the quantification of a target protein in various sample types.
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Lab products found in correlation
Sybr green master mix, by Thermo Fisher Scientific (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Superscript 2 reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Il 13, by R&D Systems (1 mentions)
2 protocols using duoset elisa kit
1
Quantifying TSLP and IgE Responses
2
Cytokine Analysis of Lung Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Lungs were prepared for cytokine analysis as previously described [52 (link),59 (link)]. Supernatants were analyzed for cytokines levels of IL-4 (eBioscience), IL-13 (R&D Systems, purified clone 38213.11 and biotinylated polyclonal goat anti-mouse IL-13), IL-17A (R&D Systems DuoSet ELISA Kit), and IFN-γ (eBioscience).
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