Determination of cytokines (IL-35, IL-10, TGF-β) was performed using the available kits – for IL-35 with the detection range of 15.6–1000 pg/ml (Wuhan Fine Biological Technology, China), for IL-10: 1.56-50 pg/ml (Diaclone, France) and for TGF-β1: 1.2–600 pg/ml (Demeditec Diagnostic GmbH, Germany), according to the manufacturer’s protocol. Validity of the assays was confirmed by parallelism between the standard curves and series of dilutions of randomly chosen plasma samples. The intra-assay coefficient of variation was < 5%. The inter-assay factor of variations was not calculated (all analyses were done in one assay). The sensitivities of the assay were 9.4, 0.98 and 1.9 pg/ml for IL-35, IL-10 and TGF-β1, respectively. We assessed sera from 31 psoriatic patients and 6 from controls.
Il 10
Manufactured by Diaclone
Sourced in France
IL-10 is a recombinant protein that functions as an anti-inflammatory cytokine. It is used in research applications to study immune system regulation and signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Ifn γ, by Diaclone (3 mentions)
Il 1β, by Diaclone (2 mentions)
Ifn γ, by Thermo Fisher Scientific (2 mentions)
Tnf α, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Interleukin 4 (il 4), by Elabscience (1 mentions)
Il 12, by Elabscience (1 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
Concanavalin a (cona), by Merck Group (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
24 well plate, by Corning (1 mentions)
Interleukin 6 (il 6), by Diaclone (1 mentions)
Interleukin 4 (il 4), by Diaclone (1 mentions)
Il 13, by R&D Systems (1 mentions)
Ip 10, by Diaclone (1 mentions)
Adiponectin, by Assaypro (1 mentions)
Konelab 20 analyzer, by Thermo Fisher Scientific (1 mentions)
11 protocols using il 10
1
Cytokine Determination in Psoriasis
2
Spleen Cell Cytokine Profiling
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Mice were sacrificed on day 35 and their spleen collected and macerated. Spleen cells (2.0 × 106) were cultured in RPMI 1640 (Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (Gibco) and antibiotic and antifungal agents (penicillin 10,000 IU/ml, streptomycin 10 mg/ml and amphotericin B 25 mg/mL) (Gibco) in 24-well plates (Corning) and incubated for 24 h at 37 °C in 5% CO2 atmosphere. Culture medium was replaced after 24 h and the cells were stimulated with 10 µg of TTFC, 10 µg of concanavalin A (ConA; Sigma-Aldrich), and with RPMI 1640, and incubated for 72 h under the same conditions. ConA and RPMI were used as positive and negative control, respectively, for cell stimuli. Supernatants were harvested from cultures and analysed by Murine ELISA kit to detected production of followed cytokines IL-4 (Elabscience, USA), IL-6 (Diaclone, France), IL-10 (Diaclone), IL-12 (Elabscience), and IFN-γ (Diaclone). The assays were performed according to the manufacturers’ instructions.
3
Quantifying Inflammatory Cytokines in Blood
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The subjects had their blood samples collected between 7 and 9 in the morning and sent for laboratory tests. The procedure involved centrifugation at 3500 rpm for 10 min and then placing the samples in 2 mL tubes for storage at −70 °C. The following markers were measured with the ELISA method: IL-4, (Diaclone SAS, Besancon, France); IL-8 (Diaclone SAS, Besancon, France); IL-10, (Diaclone SAS, Besancon, France); IL-1β, (Diaclone SAS, Besancon, France).
The manufacturer’s instructions were used to perform lab analysis. This involved checking the results and measuring concentrations. After plates had been coated with monoclonal antibodies for selected markers, the plasma samples were incubated for 60 min. Some of them bound to specific antibodies and all unbound antibodies were removed. The samples were incubated for 30 min. Afterwards, a colour reaction was induced, depending on the concentration levels.
The lower concentration levels of studied cytokines were IL-4, (0.31 pg/mL); IL-8, (29.0 pg/mL); IL-10, (4.9 pg/mL); IL-1β, (6.5 pg/m). If necessary, technical replicates were used in the study. All samples were assayed at the same time.
The manufacturer’s instructions were used to perform lab analysis. This involved checking the results and measuring concentrations. After plates had been coated with monoclonal antibodies for selected markers, the plasma samples were incubated for 60 min. Some of them bound to specific antibodies and all unbound antibodies were removed. The samples were incubated for 30 min. Afterwards, a colour reaction was induced, depending on the concentration levels.
The lower concentration levels of studied cytokines were IL-4, (0.31 pg/mL); IL-8, (29.0 pg/mL); IL-10, (4.9 pg/mL); IL-1β, (6.5 pg/m). If necessary, technical replicates were used in the study. All samples were assayed at the same time.
4
Cytokine Profiling in Co-culture
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5
Cytokine Blocking Antibodies Procurement
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Anti-human cytokine blocking Abs were purchased as follows: IFN-γ and tumor necrosis factor-alpha (TNF-α) from eBioscience Inc. (San Diego, CA), interleukin (IL)-4 from U-cytech (Utrecht, Netherland), and IL-10 from Diaclone (Besançon Cedex, France). Anti-murine IFN-γ and TNF-α blocking Abs were purchased from Thermo Scientific (Waltham, MA) and R&D Systems, respectively.
6
Cytokine Profiling via ELISA
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IP-10 (851.870.015, Diaclone Research, Besancon, France), IL-10 (851.540.015, Diaclone Research, Besancon, France) and CCL4 (DY271-05, R&D systems, Minneapolis, USA) ELISA kits were used. ELISA testing was performed according to the manufacturer’s protocol using coating antibody clones B-S10, B-C50 and #24006 and detection antibodies B-T10, BC-55 and BAF271 respectively for IL-10, IP-10 and CCL4. HRP absorbance was determined at wavelength of 450 nm.
7
Metabolic Biomarkers in Aging Rats
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Plasma levels of fasting glucose, triglycerides, and total cholesterol were determined using a Konelab 20 analyzer (Thermo-Electron Corporation, Waltham, MA, USA). ELISA kits were used to determine insulin (Alpco Diagnostics, Salem, NH, USA), leptin, (Biovendor, Bmo, Czech Republic), adiponectin (AssayPro, St Charles, MO, USA), TNFα (Millipore, Molsheim, France) and IL-10 (Diaclone, Besançon, France). Homeostatic model assessment of insulin resistance (HOMA-IR) was calculated to assess insulin sensitivity in old rats, using the formula:
with fasting glucose level expressed as mmol/L and fasting insulin level expressed as mIU/L.
with fasting glucose level expressed as mmol/L and fasting insulin level expressed as mIU/L.
8
Serum Biomarker Quantification Protocol
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9
Immunological Biomarker Assessment Protocol
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Following are
the ELISA kits used for tumor necrosis factor alpha (TNF-α),
caspase-3 (cat no: E-EL-RO160, Elabscience, Beijing, China), MCP-1
(cat no: ab2194045, Abcam, Cambridge, UK), CK-MB (cat no: abx155346,
Abcam, Cambridge, UK), BAX (cat no: LS-F35417, LSBio, WA, USA), NF-κB
(cat no: LS-F69373, LSBio Inc. WA, USA), TNF-α (cat no: 88-7340-22,
Invitrogen, Thermo Fischer, USA), IL-1β (cat no: 670.040.096,
Diaclone SAS, France), and IL-10 (cat no: 670 020, Diaclone SAS, France),
and the assay was carried out as per the instructions of the manufacturer.
the ELISA kits used for tumor necrosis factor alpha (TNF-α),
caspase-3 (cat no: E-EL-RO160, Elabscience, Beijing, China), MCP-1
(cat no: ab2194045, Abcam, Cambridge, UK), CK-MB (cat no: abx155346,
Abcam, Cambridge, UK), BAX (cat no: LS-F35417, LSBio, WA, USA), NF-κB
(cat no: LS-F69373, LSBio Inc. WA, USA), TNF-α (cat no: 88-7340-22,
Invitrogen, Thermo Fischer, USA), IL-1β (cat no: 670.040.096,
Diaclone SAS, France), and IL-10 (cat no: 670 020, Diaclone SAS, France),
and the assay was carried out as per the instructions of the manufacturer.
10
Cytokine Profiling of Restimulated Cells
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Cells were restimulated on day 7 with 20 ng/ml phorbol myristate acetate (PMA) and 0.5 μM ionomycin (Sigma Aldrich) for 6 h. The levels of IL-17A, IL-21, IL-22, IFNγ, IL-9 (eBioscience, San Diego, CA, USA), and IL-10 (Diaclone, Madrid, Spain) in supernatants were analyzed by ELISA. Final values were corrected considering the final volume of the cultures and the number of viable cells in each sample.
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