Rmil 17a

Manufactured by Thermo Fisher Scientific

RmIL-17A is a laboratory instrument designed for the detection and quantification of interleukin-17A (IL-17A) in biological samples. It utilizes a reliable and reproducible method to measure IL-17A levels, which is a key cytokine involved in inflammatory and autoimmune processes.

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4 protocols using rmil 17a

Modulating IL-17A in Allergic Asthma

Cited 1 time

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IL-17A mab (Bio X Cell: 50 μg/mouse and 100 μg/mouse) or mouse IgG isotype control antibody (50 μg/mouse and 100 μg/mouse) was administered separately via the intraperitoneal route.13 (link)19 (link) Recombinant mouse IL-17A (rmIL-17A; PeproTech) was dissolved in sterile phosphate buffered saline (PBS) and instilled at a dose of 1 μg/mouse per time.13 (link) During allergen sensitization, IL-17A mab and rmIL-17A were given every 3 days beginning from the first to the second immunization for 3 times after each sensitization in total (marked as “TDI + IL-17A mab/S” and “TDI + recombinant IL [rIL]-17A/S”), while during the TDI challenge phase, IL-17A mab and rmIL-17A were given immediately after each inhalation (marked as “TDI + IL-17A mab/C” and “TDI + rIL-17A/C”).

Chen S., Yu L., Deng Y., Liu Y., Wang L., Li D., Yang K., Liu S., Tao A, & Chen R. (2022). Early IL-17A Prevention Rather Than Late IL-17A Neutralization Attenuates Toluene Diisocyanate-Induced Mixed Granulocytic Asthma. Allergy, Asthma & Immunology Research, 14(5), 528-548.

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Intradermal Cytokine Injections for Wound Healing

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The wound edge was intradermally injected with either rm IL-17A (500 ng in 20 μl of PBS, carrier-free; PeproTech) or PBS control immediately after a 4-mm biopsy. The following cytokine concentrations were used for daily intradermal injections for 7 days in unwounded ear pinnae and compared with the PBS-only vehicle control: IL-17A (500 ng in 20 μl of PBS, carrier-free; PeproTech), IL-17F (1 μg in 20 μl of PBS, carrier-free; PeproTech), or IL-22 (500 ng in 20 μl of PBS, carrier-free; PeproTech).

Konieczny P., Xing Y., Sidhu I., Subudhi I., Mansfield K.P., Hsieh B., Biancur D.E., Larsen S.B., Cammer M., Li D., Landén N.X., Loomis C., Heguy A., Tikhonova A.N., Tsirigos A, & Naik S. (2022). Interleukin-17 governs hypoxic adaptation of injured epithelium. Science (New York, N.Y.), 377(6602), eabg9302.

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Suppression of CD8+ T Cell Proliferation by Tumor-Derived Neutrophils

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For proliferation assay, purified splenic CD8⁺ T cells from C57BL/6 mice were labeled with 2.5 uM CellTrace ef670 (eBioscience) in PBS for 6 min at 37°C. The labeling reaction was quenched by addition of cold DMEM medium with 10% FCS. Sorted CD11b+Ly6G+ cells from spleens of LLC tumor-bearing mice or isolated neutrophils from bone marrows of naive C57BL/6 mice were cocultured with polyclonal-stimulated (5 ug/mL plate-bound anti-CD3ε (2C11) and 5 ug/mL anti-CD28 (37.51)) and ef670-labeled splenic CD8⁺ T cells at a series of ratios (4:1, 1:1, 1:4) in the presence or absence of 10ng/mL of rmIL-17A (Peprotech). The proliferation and IFNγ production of CD8⁺ T cells was evaluated 3 d later by flow cytometry as described above.

Liu Y., O'Leary C.E., Wang L.C., Bhatti T.R., Dai N., Kapoor V., Liu P., Mei J., Guo L., Oliver P.M., Albelda S.M, & Worthen G.S. (2015). CD11b+Ly6G+ cells inhibit tumor growth by suppressing IL-17 production at early stages of tumorigenesis. Oncoimmunology, 5(1), e1061175.

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Dissection and Culturing of Murine Skin Organoids

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Epithelial organoids were generated as previously described from the dorsal skin of C56BL/6 mice (ages postnatal days 46 to 50) (41 (link)). One day after culturing in normoxia (21% O₂) or hypoxia (2% O₂), rmIL-17A (500 ng/ml, Peprotech), TNFα (100 ng/ml, Peprotech) (42 (link)), IL-1β (100 ng/ml, Peprotech) (43 (link)), IL-6 (100 ng/ml, Peprotech) (44 (link)), epidermal growth factor (500 ng/ml, Peprotech) (41 (link)), or PBS was added to the organoid culture medium and replenished on D3. Organoids were harvested on D6 using a nonenzymatic cultrex organoid harvesting solution (R&D Systems) for downstream analyses. For inhibitor studies, organoids were treated 12 hours before harvest with 60 nM rapamycin (LC Laboratories) (45 (link)) and vehicle control or 3 hours with U0126-ERK1/2 inhibitor (10 μM, MedChemExpress) (46 (link)) or MK-2206-AKT inhibitor (5 μM, MedChemExpress) (47 (link)) and vehicle control. Inhibitor concentrations and time course were chosen on the basis of published studies, efficacy of inhibition, and conditions that minimized organoid cell death in our cultures. For protein stability studies, cycloheximide (50 μg/ml, Cayman Chemical) was added to the culture for 5, 10, and 15 min. For mRNA stability studies, actinomycin D (10 μg/ml, Cayman Chemical) was added to the culture for 2, 5, and 8 hours.

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