Tgf β

Manufactured by BioLegend

Sourced in United States, United Kingdom

TGF-β is a multifunctional cytokine that regulates a variety of cellular processes, including cell growth, differentiation, and immune function. It is a key regulator of the immune system and plays a crucial role in the maintenance of immune homeostasis.

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Lab products found in correlation

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Close spelling variants of this product

Recombinant human TGF-β (9 citations) Anti-TGF-β (4 citations) Human-TGF-β (3 citations) TGF-β ELISA kits (3 citations) Anti-human/mouse TGF-β (19D8) (2 citations) Rat-anti-TGF-β (2 citations) Recombinant murine TGF-β (2 citations)

94 protocols using tgf β

TGF-β-Induced NFAT Activation in Jurkat Cells

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NFAT EGFP reporter Jurkat cells transduced with the TGF‐β CAR were seeded at 5–10 × 10⁴ cells/100 μL/well in triplicate in 96‐well plates, with indicated levels of human TGF‐β or human latent latency associated peptide (LAP)‐bound TGF‐β (BioLegend). Reporter induction was assessed by flow cytometry after 17 hr at 37°C.

Hou A.J., Chang Z.L., Lorenzini M.H., Zah E, & Chen Y.Y. (2018). TGF‐β–responsive CAR‐T cells promote anti‐tumor immune function. Bioengineering & Translational Medicine, 3(2), 75-86.

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Th17 and Tc17 Cell Differentiation

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CD4⁺ CD25⁻CD62L^high cells were purified from C57BL/6 splenocytes using an EasySep Mouse Naïve CD4⁺ T cell isolation kit from STEMCELL Technologies (Vancouver, Canada), and they were differentiated into Th17 cells (TGF-β, 2 ng/ml; IL-6, 20 ng/ml; Anti-IFN-γ, 10 μg/ml; Anti-IL-4, 10 μg/ml, BioLegend, San Diego, CA) in the presence of plate-bound anti-CD3 (5 μg/ml, BioLegend, San Diego, CA) and anti-CD28 (2 μg/ml, BioLegend, San Diego, CA). The cells were harvested and processed for cytokine analysis at the RNA or protein level using real-time qPCR, flow cytometry, and ELISA on day five. Alternatively, splenocytes from OT-I mice were activated using OVA-derived peptides SIINFEKL (50 ng/ml, Sangon Biotech, Shanghai, China) and polarized to Tc17 cells using cytokine TGF-β (2 ng/ml, BioLegend, San Diego, CA) and IL-6 (20 ng/ml, BioLegend, San Diego, CA) for four or five days.

Xia L., Tian E., Yu M., Liu C., Shen L., Huang Y., Wu Z., Tian J., Yu K., Wang Y., Xie Q, & Zhu D. (2022). RORγt agonist enhances anti-PD-1 therapy by promoting monocyte-derived dendritic cells through CXCL10 in cancers. Journal of Experimental & Clinical Cancer Research : CR, 41, 155.

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Naïve T-cell Polarization Assay

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Spleen from five naïve C57BL/6 mice were harvested and a single cell suspension was prepared followed by RBC lysis. Naïve T cells were negatively selected using EasySep™ Mouse T-cell Isolation Kit (Stemcell Technologies, Vancouver Canada) as per manufacturer’s instructions. Naïve T cells were then polarized in to different subsets as described previously (Noubade et al. 2011 (link); Noubade et al. 2007 (link)). Briefly, 1×10⁵ T cells were stimulated for 72h in complete RPMI media (Genesee, San Diego, CA) with plate bound anti-CD3 (5μg/ml) and soluble CD28 antibodies (2μg/ml) (Biolegend) for the T_h0 condition. In addition, the following cytokines and antibodies were added to the T_h0 conditions for polarization: T_h1 (10μg/ml anti-IL-4 (BioLegend), 4ng/ml IL-12 (Peprotech)), T_h2 (10μg/ml anti-IFN-γ (Biolegend), 30ng/ml IL-4 (Peprotech)), T_h17 (30ng/ml IL-6, 1ng/ml TGF-β (Biolegend), 10μg/ml anti-IFN-γ, 10μg/ml anti-IL-4) and T_reg (2ng/ml TGF-β). Cultures were harvested for RNA and supernatants.

Ginwala R., Bhavsar R., Moore P., Bernui M., Singh N., Bearoff F., Nagarkatti M., Khan Z.K, & Jain P. (2020). Apigenin modulates dendritic cell activities and curbs inflammation via RelB inhibition in the context of neuroinflammatory diseases. Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, 16(2), 403-424.

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Differentiation of Naive T Cells into Effector Subsets

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CD4⁺CD62L^hiFoxp3-GFP⁻ cells were sorted by fluorescence-activated cell sorting (purity >99%) and placed in culture in 24-well plates with TCRα^−/− splenocytes (1:4 T cell/splenocyte ratio), 2 µg/ml anti-CD3 (clone 2C11), 1 µM 4-hydroxytamoxifen and either 5 ng/ml TGF-β (T reg cell conditions), 5 ng/ml TGF-β + 30 ng/ml IL-6 (Th17 conditions), or 10 ng/ml IL-12 + 10 µg/ml anti-IL4 (Th1 conditions; all cytokines were obtained from BioLegend; anti-IL-4 clone 11B11 was purchased from BioXCell). Cells were harvested on day 4 of culture and analyzed by flow cytometry. For Th1 and Th17 conditions, cells were stimulated with PMA and ionomycin, as described above, before analysis.

Paterson A.M., Lovitch S.B., Sage P.T., Juneja V.R., Lee Y., Trombley J.D., Arancibia-Cárcamo C.V., Sobel R.A., Rudensky A.Y., Kuchroo V.K., Freeman G.J, & Sharpe A.H. (2015). Deletion of CTLA-4 on regulatory T cells during adulthood leads to resistance to autoimmunity. The Journal of Experimental Medicine, 212(10), 1603-1621.

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Differentiation of Naive CD4 T Cells

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Spleen and mesenteric LNs were harvested and made intro single cell suspensions. Following RBC lysis, naïve CD4 T cells were isolated according to the Mojosort kit protocol (Biolegend). Cell culture plates were coated with anti-CD3ε (5ug/ml, Biolegend, 145-2C11) and anti-CD28 (5ug/ml, Tonbo, 37.51) for 3-4 hours at 37C. Naïve CD4 T cells were plated in coated wells at .5x10^6/ml for 5 days in appropriate Th polarizing conditions (T_H0: IL-2 (50U/ml); T_H1: IL-2 (50U/mL), IL-12 (10ng/mL), anti-IL-4 (10ug/mL, 11B11); T_H2: IL-2 (50U/mL), IL-4 (4ng/mL), anti-IFNg (10ug/mL, XMG1.2); T_H17: IL-6 (20ng/mL), TGFb (5ng/mL), anti-IL-4 (10ug/mL, 11B11), anti-IFNg (10ug/mL, XMG1.2), IL-23 (20ng/mL), IL-1b (20ng/mL)). Following differentiation, cells were replated at 1x10^6/ml in the presence of IL-2 (10U/ml) and rested for 2 additional days to allow differentiation into T_EM. For generation of iTr_egs, naïve CD4 T cells were plated in anti-CD3ε (3ug/ml, 145-2C11) coated wells in the presence of soluble anti-CD28 (1ug/ml, 37.51), IL-2 (50U/ml), and TGF-b (5ng/ml, Biolegend) for 3 days.

McDaniel M.M., Chawla A.S., Jain A., Meibers H.E., Saha I., Gao Y., Jain V., Roskin K., Way S.S, & Pasare C. (2022). Effector Memory CD4+ T cells induce damaging innate inflammation and auto-immune pathology by engaging CD40 and TNFR on myeloid cells. Science immunology, 7(67), eabk0182.

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Differentiation of Induced Regulatory T Cells

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To investigate the differentiation of iTreg, CD4⁺ naïve T cells were purified using a commercially available isolation kit according to the manufacturer’s instructions (Miltenyi Biotec, Germany) and stimulated with plate-coated anti-CD3 antibody (1 mg/ml, Biolegend, USA) and soluble anti-CD28 antibody (1 mg/ml, Biolegend, USA) in X-VIVO15 medium (Lonza, Switzerland). Three days after plating, TGF-b (1 ng/ml, Biolegend, USA) and interleukin 2 (IL-2, 50 U/ml, Biolegend, USA) were added in the culture, and the cells were continued to cultivate for 2 days. According to different experimental purposes, reagents were added together with the polarization cytokines, including methyl-b-cyclodextrin-cholesterol (cholesterol, 10–20 mg/ml, Sigma, USA), dimethyloxalylglycine (DMOG, 100 mM, MCE, USA), PX-478 (10 mM, MCE, USA), MT (10 mM, Sigma, USA) and IL-1b (100 ng/ml, Biolegend, USA).

Zhang H., Xia N., Tang T., Nie S., Zha L., Zhang M., Lv B., Lu Y., Jiao J., Li J, & Cheng X. (2023). Cholesterol suppresses human iTreg differentiation and nTreg function through mitochondria-related mechanisms. Journal of Translational Medicine, 21, 224.

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Polarization of Naive CD4+ T Cells

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Naive CD4⁺ T cells were enriched from PBMCs by negative magnetic separation using MojoSort Human CD4 Naive T cell isolation Kit (BioLegend). The purity of isolated cells was checked by flow cytometry (> 95%). 2.5 × 10⁵ naive cells were cultured in flat-bottom 96-well plates with CD3/CD28 Dynabeads in RPMI 1640 supplemented with 15% FCS, 10 mM HEPES (Sigma), 100 U/ml penicillin (Sigma), and 200 mM L-glutamine (Sigma) for 4 days under the following conditions: Th0 (anti-IL-4 (5 μg/ml), anti-IFNγ (5 μg/ml)); Th1 (anti-IL-4 (5 μg/ml), IL-12 (10 ng/ml)); Th2 (IL-4 (10 ng/ml), anti-IFNγ (5 μg/ml)); Th17 (IL-6 (20 ng/ml), TGFβ (1 ng/ml), anti-IFNγ (5 μg/ml), anti-IL-4 (5 μg/ml)) (all recombinant proteins and antibodies were from BioLegend). On day 4, cells were stimulated with PMA (50 ng/ml) and ionomycin (375 ng/ml) for 5 h. Golgi Plug was added into the culture (1:1000 dilution) 1 h after stimulation. The levels of IFNγ, IL-4, and IL-17A were measured by flow cytometry with intracellular staining protocol.

Aba Ü., Maslak İ.C., İpşir C., Pehlivan D., Warnock N.I., Tumes D.J., Cildir G, & Erman B. (2024). A Novel Homozygous Germline Mutation in Transferrin Receptor 1 (TfR1) Leads to Combined Immunodeficiency and Provides New Insights into Iron-Immunity Axis. Journal of Clinical Immunology, 44(2), 55.

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T cell Differentiation Induction Protocol

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For Th0 conditions, naive CD4⁺ T cells were sorted by MACS and were stimulated for 5 d with plate-bound antibodies against CD3 (145-2C11; 2 µg/ml; eBioscience) and CD28 (PV-1; 2 µg/ml; BioLegend). For Th2 cell differentiation, 50 ng/ml IL-4 (BioLegend) and 10 µg/ml neutralizing anti–IFN-γ mAb (XMG 1.2; BioLegend) were added. For Th17 cell differentiation, 40 ng/ml IL-6 (BioLegend), 40ng/ml IL-23 (R&D Systems), 1 ng/ml TGF-β (BioLegend), 10 µg/ml anti–IL-4 (11B11; BioLegend), and 10 µg/ml anti–IFN-γ (XMG1.2; BioLegend) neutralizing antibodies were added. For Th1 cell differentiation, 5 ng/ml IL-12 (BioLegend) and 10 µg/ml anti–IL-4 neutralizing mAbs (11B11; BioLegend) were added.

Mooster J.L., Le Bras S., Massaad M.J., Jabara H., Yoon J., Galand C., Heesters B.A., Burton O.T., Mattoo H., Manis J, & Geha R.S. (2015). Defective lymphoid organogenesis underlies the immune deficiency caused by a heterozygous S32I mutation in IκBα. The Journal of Experimental Medicine, 212(2), 185-202.

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Novel Nanodiamond-based Immunomodulation

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Commercially prepared nanodiamond powder (C_ND) was purchased from Millipore Sigma, USA (Cat No. 636444). The raw diamond powder was collected from Diamond Market, Surat, India. Primary or conjugated anti-mouse CD40, CD86, CD3, CD4, CD8, CD11c and CD11b and CD25, TNF-α, TGF-β, and Foxp3 were purchased from BioLegend USA. Information about the antibodies is summarized in Supplementary Table S1 included in the supplementary information file. β-actin was purchased from Cell Signaling Technology. Annexin-FITC apoptosis assay kits and ELISA kits were procured from BioLegend USA. LPS and all other fine chemicals were purchased from Millipore Sigma, USA.

Paladhi A., Rej A., Sarkar D., Singh R., Bhattacharyya S., Sarkar P.K., Kar P.K., Manna P.P, & Hira S.K. (2022). Nanoscale Diamond-Based Formulation as an Immunomodulator and Potential Therapeutic for Lymphoma. Frontiers in Pharmacology, 13, 852065.

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Regulatory T Cell Induction Protocol

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CD4⁺CD25^– cells were isolated from the spleens of mice by MACS using EASYSEP Magnets (Stemcell Technologies), according to the manufacturer’s protocol and placed in a 96-well plate. Tregs were induced by addition of anti–mouse CD3 (clone 145-2C11, eBioscience) and CD28 (clone 37.51, eBioscience) antibodies (3 μg/mL in complete RPMI media), TGF-β (10 ng/mL, BioLegend) and IL-2 (100 ng/mL, BioLegend) cytokines, and ACTH (100ng/mL, MilliporeSigma). The plate was incubated in 5% CO₂ at 37°C for 3 days.

Zhao J., Jiang L., Uehara M., Banouni N., Al Dulaijan B.S., Azzi J., Ichimura T., Li X., Jarolim P., Fiorina P., Tullius S.G., Madsen J.C., Kasinath V, & Abdi R. (2021). ACTH treatment promotes murine cardiac allograft acceptance. JCI Insight, 6(13), e143385.

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