Rat il 10

Manufactured by R&D Systems

Sourced in United States

Rat IL-10 is a recombinant protein that represents the interleukin-10 cytokine from the rat. Interleukin-10 is an anti-inflammatory cytokine that regulates the immune response.

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IL-10 (345 citations) Rat IL-10 DuoSet ELISA (3 citations) Rat IL-10 kit (3 citations) Rat IL-10 ELISA kit (2 citations) Rat IL-10 Quantikine ELISA Kit (2 citations) Recombinant rat IL-10 (2 citations)

6 protocols using rat il 10

Liver Tissue Cytokine Extraction and Quantification

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The extraction method of liver tissues was described by Chen et al. [58 (link)]: 0.5 g of liver tissues was homogenized in 1.5 mL of ice-cold buffer containing 50 mM Tris (pH 7.2), 150 mM NaCl, 1% Triton X-100, and 0.1% protease inhibitor (PI) (HYK0010, MedChemExpress, Monmouth Junction, NJ, USA). The homogenized solution was then centrifuged at 3000 rpm for 15 min at 4 °C, and the supernatant was collected. Concentrations of hepatic TNF-α, IL-1β, IL-6, and IL-10 were determined by corresponding enzyme-linked immunosorbent assay (ELISA) kits, including rat TNF-α (BioLegend Systems, San Diego, CA, USA), IL-1β (Rat IL-1, R&D Systems, Minneapolis, MN, USA), IL-6 (Rat IL-6, R&D Systems) and IL-10 (Rat IL-10, R&D Systems, Minneapolis, MN, USA). The OD was read at 450 nm with a microplate reader (Molecular Devices, Sunnyvale, CA, USA) for all cytokines.

Chen T.Y., Chen Y.L., Chiu W.C., Yeh C.L., Tung Y.T., Shirakawa H., Liao W.T, & Yang S.C. (2022). Effects of the Water Extract of Fermented Rice Bran on Liver Damage and Intestinal Injury in Aged Rats with High-Fat Diet Feeding. Plants, 11(5), 607.

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Cytokine Profiling in Cell Culture

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Cell culture supernatants (48-hours) were analyzed in duplicate by ELISA using kits: PGE2 (multi-species), rat IL-1RA, rat IL-10 (R&D Systems, Minneapolis, MN); FGF7/KGF (BlueGene, Shanghai); MPO (RayBioTech, Norcross, GA); TSG6 (MyBiosource, San Diego, CA). Other cytokines were evaluated on a Luminex (Austin, TX) using rat-specific bead sets: FasL, G-CSF, GM-CSF, IFNγ, IL-4, IL-12p40&p70, LIX, RANTES (Millipore, Billerica, MA); PAI-1, CXCL1, TNFα, IL1α, IL-2, IL-4, IL-6 (R&D Systems).

Raza K., Larsen T., Samaratunga N., Price A.P., Meyer C., Matson A., Ehrhardt M.J., Fogas S., Tolar J., Hertz M.I, & Panoskaltsis-Mortari A. (2014). MSC Therapy Attenuates Obliterative Bronchiolitis after Murine Bone Marrow Transplant. PLoS ONE, 9(10), e109034.

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Microglial Phenotypical Changes Induced by Cytokines

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To investigate the phenotypical changes of microglia, MG d14 were pre-treated with 10 ng/mL GMCSF (R&D Systems, Minneapolis, MN, USA) for 5 days and then incubated with the following cytokines for 3 days: 10 ng/mL GMCSF (R&D systems, Minneapolis, MN, USA), 100 nM Substance P (SP, Sigma, St. Louis, MO, USA), 10 ng/mL rat IL-4 (R&D systems, Minneapolis, MN, USA), 10 ng/mL rat IL-10 (R&D systems, Minneapolis, MN, USA), 10 ng/mL TNF-α (Sigma, St. Louis, MO, USA), and 20 ng/mL IFN-γ (R&D systems, Minneapolis, MN, USA).

Kim S, & Son Y. (2021). Astrocytes Stimulate Microglial Proliferation and M2 Polarization In Vitro through Crosstalk between Astrocytes and Microglia. International Journal of Molecular Sciences, 22(16), 8800.

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Multiplex Cytokine and Chemokine Analysis

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A total of 50 μl of tissue sonicates, serum, or cell culture supernatants were used for the assays. Protein levels in serum samples were analyzed on multiplex ELISA cytokine (IFNγ, LOD: 25 pg/ml; IL-1β, LOD: 6.25 pg/ml; IL-2, 3.125 pg/ml; IL-6, LOD: 4.165 pg/ml; IL-10, LOD: 3.125 pg/ml; CXCL1, LOD: 0.781 pg/ml; TNF, LOD: 25 pg/ml) and chemokine arrays (CCL2, LOD: 1.172 pg/ml; CCL3, LOD: 0.293 pg/ml; CXCL2, LOD: 0.391 pg/ml; CCL20, LOD: 6.25 pg/ml) (Aushon, CA, USA). Chemiluminescence was quantified on a Signature-PLUS CCD (Aushon) and analyzed using PROarray Analyst Software (Aushon). Protein levels in tissue sonicates and cell culture supernatants were determined using ELISA kits for rat IL-1β (R&D Systems, Minneapolis, MN; LOD: 5 pg/ml), rat IL-10 (R&D Systems; LOD: 10 pg/ml), rat BDNF (Promega Corporation, Madison, WI; LOD: 15.6 pg/ml), and rat CCL2 (R&D Systems; LOD: 15.6 pg/ml). The assays were performed according to manufacturer's instructions. The analyte concentrations are presented as picograms per 100 μg of total protein.

Grace P.M., Fabisiak T.J., Green-Fulgham S.M., Anderson N.D., Strand K.A., Kwilasz A.J., Galer E.L., Walker F.R., Greenwood B.N., Maier S.F., Fleshner M, & Watkins L.R. (2016). Prior voluntary wheel running attenuates neuropathic pain. Pain, 157(9), 2012-2023.

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Inflammatory Response to Rapamycin

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To test the inflammatory response after rapamycin treatment, colorimetric ELISA kits were used to detect the following cytokines in serum and brain protein extract: rat IL-10, rat interferon (IFN)-γ, and rat TGF-β (all R&D Systems, USA). For each ELISA analysis, 40 μl of sample was used without dilution in accordance with the manufacturer's instructions.

Lu Q., Gao L., Huang L., Ruan L., Yang J., Huang W., Li Z., Zhang Y., Jin K, & Zhuge Q. (2014). Inhibition of mammalian target of rapamycin improves neurobehavioral deficit and modulates immune response after intracerebral hemorrhage in rat. Journal of Neuroinflammation, 11, 44.

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Formation of Crosslinked Microgels via Microfluidics

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Microgels were formed via water-in-oil emulsion using a microfluidic device with parallel flow-focusing particle generators as previously described^{[40 (link),46 ]}. The aqueous phase (1 mL) consisted of NorHA (30 mg, 3% w/v), thiol crosslinker (pentaerythritol tetrakis(mercaptoacetate), PETMA, TCI Chemicals or 1,4-dithiothreitol (DTT)) at 0.95 mol thiol per 1 mol norbornene, Irgacure-2959 (I2959, 0.05% w/v), HepSH (0.05 mol thiol per 1 mol norbornene), and (if indicated) rat IL-10 (R&D Systems, 12 μg) and was attached to an in-line 0.22 μm filter with a flow rate of 0.6 mL/hr. For visualization purposes, if indicated, fluorescein isothiocyanate-dextran (FITC-dextran, 2 MDa, 1 mg) or fluorescent peptide (GCDDD-5(6)-carboxyfluorescein, 0.05 mol thiol per 1 mol norbornene) was incorporated in the aqueous solution. The oil phase consisted of 2% w/w Span 80 in mineral oil and was filtered using a 0.45 μm filter prior to use at a flow rate of 3.5 mL/hr. Microgels were crosslinked via UV light (OmniCure Series 1500, Excelitas, 320–390 nm, 20 mW/cm²) as they flowed out of the device and into a collection vial. Microgels were washed with water and then isopropanol to remove oil, dried overnight on vacuum, and stored at −20°C.

Chen M.H., Chung J.J., Mealy J.E., Zaman S., Li E.C., Arisi M.F., Atluri P, & Burdick J.A. (2018). Injectable supramolecular hydrogel/microgel composites for therapeutic delivery. Macromolecular bioscience, 19(1), e1800248.

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