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Anti il10 clone jes3 19f1

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Anti-IL10 (clone JES3–19F1) is a monoclonal antibody that binds to the cytokine interleukin-10 (IL-10). It can be used for research purposes to study the function and regulation of IL-10 in various biological systems.

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Lab products found in correlation

Polycarbonate membrane insert, by Corning (2 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (2 mentions) Automacs, by Miltenyi Biotec (2 mentions) Recombinant il 10, by Thermo Fisher Scientific (2 mentions) Naive t cell isolation beads, by Miltenyi Biotec (2 mentions) T cell activation expansion kit, by Miltenyi Biotec (2 mentions) Microplate reader, by Bio-Rad (1 mentions) Duoset elisa kit, by R&D Systems (1 mentions)

3 protocols using anti il10 clone jes3 19f1

1

Modulation of T-cell Activation by APCs

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RosetteSep enriched APCs or skin isolated APCs were stimulated with anti-CD40/Poly I:C as described above for 1 hour. Naive T cells were sorted from an allogenic donor using naive T cell isolation beads (Miltenyi) and an autoMACs (Miltenyi). Naive T cells were labeled with CFSE (Life Technologies) according to standard protocols. T cells were then added to stimulated unwashed APCs in a 5:1 ratio and incubated for 6 days before staining for flow cytometry (Table S1). Alternatively, T cells were stimulated using the T cell activation/expansion kit containing beads coated with antibodies against CD2, CD3, and CD28 (Miltenyi). For transwell experiments, purified monocytes were cultured in the upper chamber and T cells stimulated with T cell activation/expansion kit in the lower chamber in a 1:5 ratio using a 24-well plate with polycarbonate membrane inserts (Corning). Where indicated, APCs were pre-incubated with a 5ug/ml neutralizing anti-IL10 (clone JES3–19F1, BD Biosciences) or isotype (clone R35–95, BD Biosciences) for 1 hour prior to anti-CD40/poly(I:C) stimulation. For IL-10 add-in experiments, recombinant IL-10 (Peprotech) was added directly to cultures. A dose of 250 pg/ml IL-10 was determined by titration.
Thompson E.A., Darrah P.A., Foulds K.E., Hoffer E., Caffrey-Carr A., Norenstedt S., Perbeck L., Seder R.A., Kedl R.M, & Loré K. (2019). Monocytes Acquire the Ability to Prime Tissue-Resident T Cells via IL-10-Mediated TGF-β Release. Cell reports, 28(5), 1127-1135.e4.
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2

Cytokine Profiling of T Cell Cultures

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Culture supernatants from amplified T cell libraries assay were tested for cytokine content. ELISA measurement of cytokines was performed with purified coating and biotinylated detection antibodies: anti-IFN-γ (clone 2G1) and biotin-labeled anti-human IFN-γ mAb (Thermo Scientific); anti-IL-10 (clone JES3-19F1) and biotin-labeled anti-human/viral IL-10 (BD Biosciences). Production of IL-17 and GM-CSF were measured with a DuoSet ELISA kit (R&D Systems, Minneapolis, MN). The absorbance was measured and concentrations were determined with a microplate reader and reader-associated software (Bio-Rad, Hercules, CA).
Cao Y., Amezquita R.A., Kleinstein S.H., Stathopoulos P., Nowak R.J, & O’Connor K.C. (2016). Autoreactive T cells from patients with myasthenia gravis are characterized by elevated IL-17, IFN-γ and GM-CSF and diminished IL-10 production. Journal of immunology (Baltimore, Md. : 1950), 196(5), 2075-2084.
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3

Modulation of T-cell Activation by APCs

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RosetteSep enriched APCs or skin isolated APCs were stimulated with anti-CD40/Poly I:C as described above for 1 hour. Naive T cells were sorted from an allogenic donor using naive T cell isolation beads (Miltenyi) and an autoMACs (Miltenyi). Naive T cells were labeled with CFSE (Life Technologies) according to standard protocols. T cells were then added to stimulated unwashed APCs in a 5:1 ratio and incubated for 6 days before staining for flow cytometry (Table S1). Alternatively, T cells were stimulated using the T cell activation/expansion kit containing beads coated with antibodies against CD2, CD3, and CD28 (Miltenyi). For transwell experiments, purified monocytes were cultured in the upper chamber and T cells stimulated with T cell activation/expansion kit in the lower chamber in a 1:5 ratio using a 24-well plate with polycarbonate membrane inserts (Corning). Where indicated, APCs were pre-incubated with a 5ug/ml neutralizing anti-IL10 (clone JES3–19F1, BD Biosciences) or isotype (clone R35–95, BD Biosciences) for 1 hour prior to anti-CD40/poly(I:C) stimulation. For IL-10 add-in experiments, recombinant IL-10 (Peprotech) was added directly to cultures. A dose of 250 pg/ml IL-10 was determined by titration.
Thompson E.A., Darrah P.A., Foulds K.E., Hoffer E., Caffrey-Carr A., Norenstedt S., Perbeck L., Seder R.A., Kedl R.M, & Loré K. (2019). Monocytes Acquire the Ability to Prime Tissue-Resident T Cells via IL-10-Mediated TGF-β Release. Cell reports, 28(5), 1127-1135.e4.
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