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Rmil 25

Manufactured by Johnson & Johnson
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RmIL-25 is a laboratory equipment product. It is designed for the detection and quantification of Interleukin-25 (IL-25) in various biological samples. The RmIL-25 product utilizes an enzyme-linked immunosorbent assay (ELISA) method to measure IL-25 levels. The product is intended for research use only and not for diagnostic or therapeutic purposes.

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Lab products found in correlation

Rmil 2, by BioLegend (3 mentions) Endotoxin free pbs, by Merck Group (2 mentions) Rmear11, by R&D Systems (2 mentions) Ultrapure lps eb from escherichia coli 0111 b4, by InvivoGen (2 mentions) Rmil 33, by Thermo Fisher Scientific (2 mentions) Cxcl1, by R&D Systems (2 mentions) Rmil 7, by BioLegend (2 mentions) Control rigg1, by BioLegend (2 mentions) Anti il 13, by Thermo Fisher Scientific (2 mentions) Soluble anti cd28, by BioXCell (2 mentions) Cell stimulation cocktail, by Thermo Fisher Scientific (2 mentions) Anti il 4, by BioLegend (2 mentions) Celltrace violet cell proliferation kit, by Thermo Fisher Scientific (2 mentions) Anti cd3ε, by BioLegend (2 mentions) Matrigel, by Thermo Fisher Scientific (1 mentions) Collagenase 1, by Thermo Fisher Scientific (1 mentions) Matrigel, by Corning (1 mentions) Dnase 1, by Roche (1 mentions)

5 protocols using rmil 25

1

Adipose-derived MSC and ILC2 Interaction

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WAT-MSCs were purified from naive BALB/c mice. ILC2s were purified from WT BALB/c or Il4−/−Il13−/− BALB/c mice injected intraperitoneally with IL-25 and IL-2/anti–IL-2 complex (rmIL-2; 0.5 µg/mouse; BioLegend) and anti–IL-2 JES6-1A12 (0.25 µg/mouse; 2BScientific) preincubated at 37°C to form a complex and rmIL-25 (1 µg/mouse; Janssen) on 3 consecutive d. Isolated WAT-MSCs and ILC2s were co-cultured (MSCs + WT ILC2 or MSCs + IL-13/IL-4–deficient ILC2) at a ratio of 1:1 (100,000 of each) overnight in complete RPMI with IL-7 (10 ng/ml). The combined cells were then resuspended at 2 × 105 per 100 µl of Matrigel (Phenol Red-Free Corning), and injected subcutaneously into either the left or right inguinal fat pad of the same mouse. Matrigel plugs were dissected 48 h later, mechanically dissociated in RPMI-1640, digested with collagenase I (Life Technologies) and DNase I (Roche) at 37°C while shaking, and passed through a filter to obtain a single-cell suspension for analysis by flow cytometry.
Rana B.M., Jou E., Barlow J.L., Rodriguez-Rodriguez N., Walker J.A., Knox C., Jolin H.E., Hardman C.S., Sivasubramaniam M., Szeto A., Cohen E.S., Scott I.C., Sleeman M.A., Chidomere C.I., Cruz Migoni S., Caamano J., Jorgensen H.F., Carobbio S., Vidal-Puig A, & McKenzie A.N. (2019). A stromal cell niche sustains ILC2-mediated type-2 conditioning in adipose tissue. The Journal of Experimental Medicine, 216(9), 1999-2009.
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2

Investigating Cytokine and Allergen Effects

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In all instances recombinant proteins, extracts, and LPS were given in endotoxin-free PBS (Sigma-Aldrich) or PBS as a control.
rmIL-25 (2 μg/dose, made in-house, Janssen Pharmaceuticals), rmIL-33 (0.5 μg/dose, Peprotech) or rmEar11 (2 μg/dose) were given on 2 or 3 consecutive days (as indicated) and tissues harvested 24 h after the final dose. rmEar11 (2 μg/dose) or CXCL1 (0.5 μg/ml, R and D Systems; <100 EU/mg protein) were given as a single dose and tissues were harvested at 12 h post injection. rmEar11 and CXCL1 were boiled for 60 min to denature the protein.
RWP (100–200 μg/dose, RWP, Ambrosia artemisiifolia, short form, Greer Laboratories) was given intranasally on 5 consecutive days. All tissues were harvested 24 h after the final dose. Alternatively, three doses RWP were administered intranasally over 3 days and lung CD45+CD11c+F4/80+SiglecF+ alveolar macrophages purified 24 h after the final dose. Macrophage gene expression was analysed by qPCR.
LPS (1–2 μg/dose, Ultrapure LPS-EB from Escherichia coli 0111: B4, InvivoGen) was given intranasally on three consecutive days. Analyses were performed 24 h after the final dose. In some experiments, lung CD45+CD11c+F4/80+SiglecF+ alveolar macrophages were purified and gene expression analysed by qPCR.
Panova V., Gogoi M., Rodriguez-Rodriguez N., Sivasubramaniam M., Jolin H.E., Heycock M.W., Walker J.A., Rana B.M., Drynan L.F., Hodskinson M., Pannell R., King G., Wing M., Easton A.J., Oedekoven C.A., Kent D.G., Fallon P.G., Barlow J.L, & McKenzie A.N. (2020). Group-2 innate lymphoid cell-dependent regulation of tissue neutrophil migration by alternatively activated macrophage-secreted Ear11. Mucosal Immunology, 14(1), 26-37.
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3

Intranasal Administration of Inflammatory Agents

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In all instances recombinant proteins, extracts, and LPS were given in endotoxin-free PBS (Sigma-Aldrich) or PBS as a control.
rmIL-25 (2 μg/dose, made in-house, Janssen Pharmaceuticals), rmIL-33 (0.5 μg/dose, Peprotech) or rmEar11 (2 μg/dose) were given on 2 or 3 consecutive days (as indicated) and tissues harvested 24 h after the final dose. rmEar11 (2 μg/dose) or CXCL1 (0.5 μg/ml, R and D Systems; <100 EU/mg protein) were given as a single dose and tissues were harvested at 12 h post injection. rmEar11 and CXCL1 were boiled for 60 min to denature the protein.
RWP (100–200 μg/dose, RWP, Ambrosia artemisiifolia, short form, Greer Laboratories) was given intranasally on 5 consecutive days. All tissues were harvested 24 h after the final dose. Alternatively, three doses RWP were administered intranasally over 3 days and lung CD45+CD11c+F4/80+SiglecF+ alveolar macrophages purified 24 h after the final dose. Macrophage gene expression was analysed by qPCR.
LPS (1–2 μg/dose, Ultrapure LPS-EB from Escherichiacoli 0111:B4, InvivoGen) was given intranasally on three consecutive days. Analyses were performed 24 h after the final dose. In some experiments, lung CD45+CD11c+F4/80+SiglecF+ alveolar macrophages were purified and gene expression analysed by qPCR.
Panova V., Gogoi M., Rodriguez-Rodriguez N., Sivasubramaniam M., Jolin H.E., Heycock M.W., Walker J.A., Rana B.M., Drynan L.F., Hodskinson M., Pannell R., King G., Wing M., Easton A.J., Oedekoven C.A., Kent D.G., Fallon P.G., Barlow J.L, & McKenzie A.N. (2020). Group-2 innate lymphoid cell-dependent regulation of tissue neutrophil migration by alternatively activated macrophage-secreted Ear11. Mucosal Immunology, 14(1), 26-37.
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4

ILC2-Mediated Modulation of Tumor M-MDSCs

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Intestinal ILC2s were sorted and cultured in complete RPMI (RPMI-1640 + 10% FCS + penicillin/streptomycin + 2-mercaptoethanol) with 10 ng/mL rm-IL-2 (Biolegend, 575406), 10 ng/mL rm-IL-7 (Biolegend, 577806), and 20 ng/mL rm-IL-25 (Janssen) for 3 days at 37°C, and supernatant collected (ILC2-SNT). ILC2-SNT were incubated with anti-IL-4 (Biolegend, 504122) and anti-IL-13 (eBioscience, 16-7135-85) neutralizing antibodies (αIL-4/13Ab-ILC2-SNT), or control rIgG1 (Biolegend, 400432 and eBioscience, 16-4301-85) antibodies (conAb-ILC2-SNT) for 1 hour on ice. Tumor M-MDSCs were sorted and incubated with αIL-4/13Ab-ILC2-SNT or conAb-ILC2-SNT for 2 hours at 37°C. Subsequently, sorted splenic CD8+ T cells (Live CD45+ TCRγδ-CD4-CD8+) stained with the CellTrace Violet Cell Proliferation Kit (ThermoFisher, C34557), were added to the ILC2-SNT-M-MDSC culture, and stimulated for 3 days with plate-coated anti-CD3ε (Biolegend, 100360) and 2 μg/mL soluble anti-CD28 (Bio X Cell, BE0015-1) at 37°C. Cell Stimulation Cocktail (eBioscience, 00-4975-93) was added for the last 4 hours of culture, before antibody staining. For Arginase 1 expression analysis in ILC2-SNT-treated M-MDSCs, tumor or splenic M-MDSCs were sorted and cultured with αIL-4/13Ab-ILC2-SNT or conAb-ILC2-SNT for 2 days, and stained with antibodies for flow cytometry.
Jou E., Rodriguez-Rodriguez N., Ferreira A.C., Jolin H.E., Clark P.A., Sawmynaden K., Ko M., Murphy J.E., Mannion J., Ward C., Matthews D.J., Buczacki S.J, & McKenzie A.N. (2022). An innate IL-25-ILC2-MDSC axis creates a cancer-permissive microenvironment for Apc-mutation-driven intestinal tumorigenesis. Science immunology, 7(72), eabn0175.
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5

ILC2-Mediated Modulation of Tumor M-MDSCs

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Intestinal ILC2s were sorted and cultured in complete RPMI (RPMI-1640 + 10% FCS + penicillin/streptomycin + 2-mercaptoethanol) with 10 ng/mL rm-IL-2 (Biolegend, 575406), 10 ng/mL rm-IL-7 (Biolegend, 577806), and 20 ng/mL rm-IL-25 (Janssen) for 3 days at 37°C, and supernatant collected (ILC2-SNT). ILC2-SNT were incubated with anti-IL-4 (Biolegend, 504122) and anti-IL-13 (eBioscience, 16-7135-85) neutralizing antibodies (αIL-4/13Ab-ILC2-SNT), or control rIgG1 (Biolegend, 400432 and eBioscience, 16-4301-85) antibodies (conAb-ILC2-SNT) for 1 hour on ice. Tumor M-MDSCs were sorted and incubated with αIL-4/13Ab-ILC2-SNT or conAb-ILC2-SNT for 2 hours at 37°C. Subsequently, sorted splenic CD8+ T cells (Live CD45+ TCRγδ-CD4-CD8+) stained with the CellTrace Violet Cell Proliferation Kit (ThermoFisher, C34557), were added to the ILC2-SNT-M-MDSC culture, and stimulated for 3 days with plate-coated anti-CD3ε (Biolegend, 100360) and 2 μg/mL soluble anti-CD28 (Bio X Cell, BE0015-1) at 37°C. Cell Stimulation Cocktail (eBioscience, 00-4975-93) was added for the last 4 hours of culture, before antibody staining. For Arginase 1 expression analysis in ILC2-SNT-treated M-MDSCs, tumor or splenic M-MDSCs were sorted and cultured with αIL-4/13Ab-ILC2-SNT or conAb-ILC2-SNT for 2 days, and stained with antibodies for flow cytometry.
Jou E., Rodriguez-Rodriguez N., Ferreira A.C., Jolin H.E., Clark P.A., Sawmynaden K., Ko M., Murphy J.E., Mannion J., Ward C., Matthews D.J., Buczacki S.J, & McKenzie A.N. (2022). An innate IL-25-ILC2-MDSC axis creates a cancer-permissive microenvironment for Apc-mutation-driven intestinal tumorigenesis. Science immunology, 7(72), eabn0175.
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