Recombinant human il 10

Manufactured by BioLegend

Sourced in United States

Recombinant human IL-10 is a cytokine that plays a key role in the regulation of immune and inflammatory responses. It is produced by a variety of cell types, including T cells, B cells, and monocytes.

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Lab products found in correlation

Succinate, by Merck Group (2 mentions) Anti il 10, by BioLegend (2 mentions) Anti sucnr1 gpr91, by Novus Biologicals (2 mentions) Flex set kit, by BD (2 mentions) Zeba spin desalting column, by Thermo Fisher Scientific (2 mentions) Annexin 5, by BD (1 mentions) Facscanto 2 multicolor flow cytometer, by BD (1 mentions) Diva software, by BD (1 mentions) Annexin 5 and 7 aad, by BD (1 mentions) Recombinant human interleukin il 4, by R&D Systems (1 mentions) Recombinant human il 6, by BioLegend (1 mentions) Recombinant human interferon ifn γ, by Thermo Fisher Scientific (1 mentions) Pan monocyte isolation kit, by Miltenyi Biotec (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Penicillin streptomycin, by Fujifilm (1 mentions) Rpmi 1640, by Fujifilm (1 mentions) Macrophage colony stimulating factor m csf, by BioLegend (1 mentions) Bio plex assay, by Bio-Rad (1 mentions) Bio plex 200 system, by Bio-Rad (1 mentions)

Spelling variants of this product

IL-10 (167 citations) Human IL-10 (7 citations) Recombinant IL-10 (6 citations)

7 protocols using recombinant human il 10

U937 Macrophage Differentiation and Activation

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For complete differentiation of U937 cells into macrophages,
1 × 10⁶ U937 cells were plated into 12-well tissue
culture plates containing 2 ml of RPMI-S at a final concentration of
200 nM of Phorbol Myristate Acetate (PMA) and incubated for 3
d.²² Adherent cells were washed three times with PBS.
These cells will be referred from now as M0 macrophages.
Activation of the macrophages was achieved by incubating M0 macrophages
previously differentiated in the presence of 100 ng/ml of LPS for 24
h.²³ After this, the medium containing LPS was
removed completely, and cells were washed with PBS. These cells are
referred to as M1 macrophages.
Likewise, M0 macrophages were treated with 20 ng/ml of IL-10 (recombinant
human IL-10, BioLegend, San Diego, CA, USA) for 24 h, after that the
cells were washed. These cells are referred to as M2 macrophages.
All incubations were at 37°C in a humidified atmosphere with 5% of
CO₂.

Sánchez-Reyes K., Pedraza-Brindis E.J., Hernández-Flores G., Bravo-Cuellar A., López-López B.A., Rosas-González V.C, & Ortiz-Lazareno P.C. (2019). The supernatant of cervical carcinoma cells lines induces a decrease in phosphorylation of STAT-1 and NF-κB transcription factors associated with changes in profiles of cytokines and growth factors in macrophages derived from U937 cells. Innate Immunity, 25(6), 344-355.

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Assessing Staphylococcus aureus Intracellular Survival

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HMDMs were treated with recombinant human IL-10 (100 ng/mL; BioLegend) or untreated for 16 h, then inoculated with WT and/or the indicated mutant S. aureus strains at an MOI of 10:1 (bacteria:macrophage) using a mono-infection or co-infection paradigm. After a 30-min phagocytosis period, HMDMs were treated with a high dose of gentamicin (100 µg/mL) for 30 min to kill remaining extracellular bacteria and then transitioned to a low gentamicin dose (1 µg/mL) to prevent intracellular accumulation of gentamicin that may occur with extended incubation periods. At the indicated time points, HMDMs were washed with phosphate-buffered saline (PBS), lysed using sterile water, and plated on trypticase soy agar (TSA) with 5% sheep blood to quantify intracellular bacterial burden. For co-infection experiments, dual plating on TSA ± erythromycin (5 µg/mL) was used since all S. aureus transposon mutants harbored an erythromycin resistance marker.

Bertrand B.P., Shinde D., Thomas V.C., Whiteley M., Ibberson C.B, & Kielian T. (2024). Metabolic diversity of human macrophages: potential influence on Staphylococcus aureus intracellular survival. Infection and Immunity, 92(2), e00474-23.

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B/T Cell Co-culture Assay for Immunoglobulin Production

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For B/T cells co-cultures, naïve or memory B cells were co-cultured with CD4⁺ T cells (2 × 10⁴ B cells - 2 × 10⁴ CD4 T cells) in the presence of endotoxin-reduced SEB (500 ng/ml; Sigma Aldrich) in cRPMI supplemented with 10% heat-inactivated FBS. Where described, anti-IL10 (10 μg/ml - clone JES3–9D7 - Biolegend LEAF™ purified antibody), anti-SUCNR1/GPR91 (20 μg/ml – Novus Biological), anti- IL21R (10 μg/ml - clone 17A12 - Biolegend LEAF™ purified antibody) or succinate (2 mM – Sigma-Aldrich) were added during the co-cultures. Sodium azide and other preservatives were removed from the antibody preparations by protein desalting with Zeba Spin Desalting Columns (7K MWCO; Thermo Fisher). IgM and IgG concentrations were measured at day 12 in the corresponding supernatants with Flex Set Kit (BD Biosciences). For T cell-independent B cell differentiation, naïve B cells (5 × 10⁴ cells) were co-cultured with irradiated (77Gy) human-CD40L transfected fibroblasts^{43 (link)} (0.5 × 10⁴ cells) in cRPMI supplemented with 10% heat-inactivated FBS. Recombinant human IL10 (Biolegend, 500 ng/ml) and/or succinate (2 mM – Sigma-Aldrich) were added during the co-cultures. IgG and IgM concentrations were measured at day 12 in the corresponding supernatants with Flex Set Kit (BD Biosciences).

Caielli S., Troggian Veiga D., Balasubramanian P., Athale S., Domic B., Murat E., Banchereau R., Xu Z., Chandra M., Chung C.H., Walters L., Baisch J., Wright T., Punaro M., Nassi L., Stewart K., Fuller J., Ucar D., Ueno H., Zhou J., Banchereau J, & Pascual V. (2018). A CD4+ T cell population expanded in Lupus blood provides B cell help through IL10 and succinate. Nature medicine, 25(1), 75-81.

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Measuring IL-10 Effects on Hepatitis B Antigen-Induced Apoptosis

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To measure the effect of IL-10 on Ag-induced apoptosis, peripheral blood mononuclear cells (PBMC) from five HLA-A201⁺ patients with acute self-limited hepatitis B (concentration of 2×10⁶/ml) were incubated for 1 h at 37 °C with 20 μg/ml of anti-IL-10R Ab (BD Pharmingen), 200 ng/ml of recombinant human IL-10 (BioLegend) or left untreated prior to the addition of Core 18-27 peptide (1 µg/ml) and human IL-2 (100 IU/ml). After a 5 h incubation, cells were extensively washed, stained with Core 18-27 dextramer, anti-CD8 and anti-CD3 mouse Abs for 15 min in the dark, then stained with Annexin V and 7AAD (BD Pharmingen), according to the Annexin V staining protocol (BD Pharmingen). The cells were acquired immediately on a FACSCANTO II multicolor flow cytometer and were analyzed with the DIVA software (BD Biosciences, Immunocytometry Systems, CA, USA).

Fioravanti J., Di Lucia P., Magini D., Moalli F., Boni C., Benechet A.P., Fumagalli V., Inverso D., Vecchi A., Fiocchi A., Wieland S., Purcell R., Ferrari C., Chisari F.V., Guidotti L.G, & Iannacone M. (2017). Effector CD8+ T cell-derived interleukin-10 enhances acute liver immunopathology. Journal of Hepatology, 67(3), 543-548.

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Macrophage Differentiation and Stimulation

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PBMCs obtained as described above were separated using MACS microbeads (Pan Monocyte Isolation Kit; Miltenyi Biotec, Bergisch Gladbach, Germany), according to the manufacturer’s protocols. Based on morphology, the monocyte purity varied in the range of 95–98 %. Monocyte-derived macrophages were obtained by culturing monocytes in RPMI 1640 (FUJIFILM Wako Pure Chemical Co.) supplemented with 10 % FBS (Sigma-Aldrich), penicillin/streptomycin (FUJIFILM Wako Pure Chemical Co.), and 25 ng/mL macrophage colony-stimulating factor (M-CSF) (BioLegend, San Diego, CA, USA) for 5 days. M1 macrophages were generated using 20 ng/mL recombinant human interferon (IFN)-γ (PeproTech, Rocky Hill, NJ, USA) and 100 ng/mL lipopolysaccharides (LPS) (FUJIFILM Wako Pure Chemical Co.) for 24 h. M2 macrophages were generated using 20 ng/mL recombinant human interleukin (IL)-4 (R&D Systems Inc.) or 20 ng/ml recombinant human IL-10 (BioLegend) for 24 h. M0 macrophages were cultured in the medium and left untreated. Macrophages were stimulated with or without 20 ng/ml recombinant human IL-6 (BioLegend), 200 ng/ml anti-human IL-6R alpha antibody (mouse monoclonal, clone #17506, R&D Systems Inc.), or 200 ng/ml mouse IgG1 isotype control (mouse monoclonal, clone #11711, R&D Systems Inc.).

Watanabe H., Mokuda S., Tokunaga T., Kohno H., Ishitoku M., Araki K., Sugimoto T., Yoshida Y., Yamamoto T., Matsumoto M., Masumoto J., Hirata S, & Sugiyama E. (2023). Expression of factor XIII originating from synovial fibroblasts and macrophages induced by interleukin-6 signaling. Inflammation and Regeneration, 43, 2.

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Quantification of Cytokines and BT-063 Production

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To quantify cytokines, BioRad's Bio-Plex assays were used and run according to the manufacturer's instructions and run on a Bio-Plex 200 System (BioRad). To quantify BT-063 production from aIL-10 GEMs, an enzymelinked immunosorbent assay (ELISA) was developed. A standard ELISA plate was coated with 2 ug/mL of recombinant human IL-10 (BioLegend, 571006). BT-063 and recombinant rat anti-human IL-10 (for the standard curve) were added to the IL-10 coated plate. HRP-conjugated mouse anti-rat IgG (Jackson IR, 212-0350168) was used to detect the rat anti-human IL-10 antibody and HRP-conjugated rat anti-human IgG (Immuno Reagents, RbxHu-003-DHRPX) was used to detect BT-063. All samples and standards were run in duplicate.

Labadie K.P., Kreuser S.A., Brempelis K.J., Daniel S.K., Jiang X., Sullivan K.M., Utria A.F., Kenerson H.L., Kim T.S., Crane C.A, & Pillarisetty V.G. (2023). Production of an interleukin-10 blocking antibody by genetically engineered macrophages increases cancer cell death in human gastrointestinal tumor slice cultures. Cancer gene therapy, 30(9).

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B/T Cell Co-culture Assay for Immunoglobulin Production

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