Rhil 4

Manufactured by BioLegend

Sourced in United States

Recombinant human Interleukin-4 (rhIL-4) is a cytokine that plays a key role in the immune system. It is involved in the regulation of various cellular processes, including inflammation and the differentiation of B cells and T cells. rhIL-4 is used as a research reagent in experimental studies to investigate its biological functions.

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Lab products found in correlation

Isolation kits, by Miltenyi Biotec (1 mentions) Rhil 2, by R&D Systems (1 mentions) αcd28, by BD (1 mentions) Nω nitro l arginine methyl ester, by Merck Group (1 mentions) Ifn γ, by Thermo Fisher Scientific (1 mentions) M csf, by BioLegend (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rhgm csf, by BioLegend (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Lymphoprep, by STEMCELL (1 mentions)

4 protocols using rhil 4

Isolation and Characterization of Immature Dendritic Cells

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Buffy coats from the venous blood of normal healthy volunteers were obtained by the Blood Transfusion Centre of Slovenia, in accordance with institutional guidelines. Peripheral blood mononuclear cells (PBMCs) were then isolated using Lymphoprep (Stemcell Technologies, Vancouver, Canada). The cells were washed twice with phosphatebuffered saline (PBS), counted, and used as a source for the immunomagnetic negative isolation of CD14-positive cells (EasySep, Stemcell Technologies, Vancouver, Canada). CD14-positive cells (the purity of CD14 + cells was always greater than 95%, as determined by flow cytometry) were cultured in an RPMI 1640 (Sigma-Aldrich, MO, USA) medium, supplemented with 10% fetal bovine serum (Gibco, NY, USA), 50 U/mL penicillin and 50 µg/mL streptomycin (Sigma-Aldrich, MO, USA), 800 U/mL of rhGM-CSF and 1000 U/mL of rhIL-4 (Biolegend, San Diego, CA) at 1 × 10 6 cells/mL. On day 3, half of the medium was exchanged with the addition of starting quantities of rhGMCSF and rhIL-4. After 5 days, non-adherent, immature DCs were harvested and characterized, by means of flow cytometry, as CD14neg, DC-SIGNhigh, CD80low, CD83high, CD86low, HLA-DRlow, CD85khigh, CD207alow.

Biagiotti G., Purić E., Urbančič I., Krišelj A., Weiss M., Mravljak J., Gellini C., Lay L., Chiodo F., Anderluh M., Cicchi S, & Richichi B. (2021). Combining cross-coupling reaction and Knoevenagel condensation in the synthesis of glyco-BODIPY probes for DC-SIGN super-resolution bioimaging. Bioorganic chemistry, 109.

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Modulating immune responses with pharmacological agents

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Where indicated, cultures were treated with the following drugs at the indicated final concentrations: 8 µg/ml anti-CD40L (Ruplizumab, Creative Biolabs) or 8 µg/ml UltraLEAF human IgG1 isotype control (clone QA16A12, BioLegend); 0.5 µg/ml trimerized rhCD40L (BioLegend), 20 ng/ml rhIL-4 (BioLegend) or PBS solvent control; 2 µM JAK inhibitors Tofacitinib or Baricitinib (both: abcr GmbH) or DMSO solvent control; 14.8 µg/ml IL6R blocking antibody Tocilizumab or human IgG1 as isotype control; 0.5 µg/ml (3.3 nM; “low” concentration) or 10 µg/ml (67 nM; “high” concentration) of each TNF blocking reagents Etanercept or Adalimumab, or human IgG1 as isotype control. Further details on plate positioning and drug treatment are provided in Sections 1 and 2 of the Supplemental materials and methods.

Schmidt A., Huber J.E., Sercan Alp Ö., Gürkov R., Reichel C.A., Herrmann M., Keppler O.T., Leeuw T, & Baumjohann D. (2020). Complex human adenoid tissue-based ex vivo culture systems reveal anti-inflammatory drug effects on germinal center T and B cells. EBioMedicine, 53, 102684.

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Isolation and Culture of Naive CD4+ T Cells

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Mediastinal lymph nodes associated with healthy donor lungs were dissected digested with 400 U/mL Collagenase Type 4, 0.1% Dispase II and 20 μg/mL DNase I for 30 min at 37°C. Digested samples were mechanically disrupted by pushing through syringe with 18-gauze needle and passage through 100 μm and 40 μm wire meshes. Naïve CD4⁺ T cells were enriched by negative selection using isolation kits (Miltenyi Biotec, 130-094-131). Purified cells (2×10⁶ per well) were cultured in a 12-well plate pre-coated with 2 μg/mL αCD3 (eBioscience, 16-0037-85). RPMI-1640 medium was supplemented with 10% FBS, 100 U/mL penicillin, 0.1 mg/mL streptomycin, 2 mM 2-mercaptoethanol. For Th0 condition, cells were cultured at the presence of 2 μg/mL αCD28 (BD Biosciences, 555725). For Th2 (Th0+IL-4) condition, cells were cultured at the presence of 2 μg/mL αCD28, 5 ng/mL rhIL-2 (R&D Systems, 202-IL-010) and 10 ng/mL rhIL-4 (BioLegend, 204-IL-010).

Wang W., Cohen J.A., Wallrapp A., Trieu K.G., Barrios J., Shao F., Krishnamoorthy N., Kuchroo V.K., Jones M.R., Fine A., Bai Y, & Ai X. (2019). Age-related dopaminergic innervation augments T helper 2-type allergic inflammation in the postnatal lung. Immunity, 51(6), 1102-1118.e7.

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Nitric Oxide Modulation of Macrophage Polarization

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PBMCs were incubated with LPS 10 μg/mL (Escherichia coli lipopolysaccharide, Sigma) for 60 min with or without 10 min pre-incubation with the nitric oxide synthase inhibitor L-NAME 10 mM (Nω-nitro-L-arginine methyl ester, Sigma) at 37 °C, 5% CO₂. Cellular levels of nitric oxide (NO) were assessed an NO probe DAF-2DA 2 μM (4,5-diaminofluorescein diacetate, Calbiochem) by flow cytometry. Briefly, cells were incubated with DAF-2DA at 37 °C for 20 min and were subsequently surface stained for CD14 (PerCP) and HLA-DR (APC) on ice. For macrophage polarization, purified CD14+ monocytes were plated in RPMI 1640 with 10% human AB serum containing with 100 ng/mL M-CSF (macrophage colony-stimulating factor, Biolegend) for 7 days for M0 differentiation. After 7 days, M1 and M2 polarization was induced by either 100 ng/mL LPS (Sigma) plus 20 ng/mL IFNγ (PeproTech), or rhIL-4 20 ng/mL (BioLegend) for 2 additional days. Cultured macrophages were stained for CD86, HLA-DR, and CD206 for analysis by flow cytometry.

Tanoue S., Chang L.Y., Li Y, & Kaplan D.E. (2015). Monocyte-derived dendritic cells from cirrhotic patients retain similar capacity for maturation/activation and antigen presentation as those from healthy subjects. Cellular immunology, 295(1), 36-45.

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