Recombinant human tgf β1

Manufactured by Proteintech

Sourced in United States

Recombinant human TGF-β1 is a protein product produced using recombinant DNA technology. TGF-β1 is a signaling molecule that plays a role in various cellular processes, including cell growth, differentiation, and immune function.

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7 protocols using recombinant human tgf β1

Regulation of T-cell Immune Responses by TGF-β

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Recombinant human TGF-β1 was purchased from Proteintech Group (Chicago, IL, USA). Recombinant murine TGF-β1 was purchased from Novoprotein (Shanghai, China). PD-1, NFATc1, lamin B, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibodies and anti-mouse/rabbit secondary antibodies for Western blot were purchased from Proteintech Group. Phosphorylated-NFATc1 (p-NFATc1) antibody for Western blot was purchased from Bioss (Beijing, China). CTLA-4 antibody for Western blot and NFATc1 antibody for chromatin immunoprecipitation (ChIP) assay were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Fluorescence-conjugated monoclonal antibodies (mAbs) to PD-1 and CTLA-4 were purchased from Bioss. Fluorescence-conjugated mAbs to CD3 and CD8 were purchased from BD Biosciences (San Jose, CA, USA). The TGF-β receptor (TGF-βR) inhibitor SB431542 and CaN inhibitor cyclosporin A (CsA) were purchased from TargetMol (Shanghai, China).

Bao S., Jiang X., Jin S., Tu P, & Lu J. (2021). TGF-β1 Induces Immune Escape by Enhancing PD-1 and CTLA-4 Expression on T Lymphocytes in Hepatocellular Carcinoma. Frontiers in Oncology, 11, 694145.

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Antibody Sources and Plasmid Construction

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Antibodies were obtained from multiple sources: anti-glyceraldehyde 3-phosphate dehydrogenase (anti-GAPDH) and anti-AP1 from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-MBD2, anti-collagen I, anti-collagen IV, anti-TGF-β, anti-ERK1/2, anti-p-ERK1/2, and anti-fibronectin from Abcam (Cambridge, MA, USA); and anti-EGR1, anti-SMAD3, and anti-p-SMAD3 from Proteintech Group (Rosemont, IL, USA),NCM Universal Antibody Diluent (New Cell and Molecular Biotech,Suzhou, China). The recombinant human TGF-β1 was obtained from Proteintech Group. The plasmids containing the methylation promoter of EGR1 CpG-free pCpGI luciferase reporter, MBD2 and mtMBD2 (the deletion of the methylated DNA binding domain), were constructed by the Ruqi Biology (Guangdong, Guangzhou, China), according to previously published reports.²⁸ (link)^,³⁹ (link)

Ai K., Li X., Zhang P., Pan J., Li H., He Z., Zhang H., Yi L., Kang Y., Wang Y., Chen J., Li Y., Xiang X., Chai X, & Zhang D. (2022). Genetic or siRNA inhibition of MBD2 attenuates the UUO- and I/R-induced renal fibrosis via downregulation of EGR1. Molecular Therapy. Nucleic Acids, 28, 77-86.

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Antibody and Plasmid Sources for Cell Studies

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Antibodies were obtained from multiple sources: anti-GAPDH from Santa Cruz Biotechnology (Santa Cruz, CA, USA), anti-MBD2, anti-collagen I, anti-α-SMA and anti-fibronectin from Abcam (Cambridge, MA, USA), and anti-F4/80, anti-collagen IV, and anti-G0S2 from Proteintech Group (Rosemont, IL, USA). The recombinant human TGF-β1 was obtained from Proteintech Group. The plasmids containing the methylation promoter of the G0S2 CpG-free pCpGI luciferase reporter, MBD2, and mtMBD2 (the deletion of the methylated DNA binding domain), were constructed by the Ruqi Biology Company (Guangdong, Guangzhou, China) according to previously published reports [15 (link), 17 (link)]. The MBD2 siRNA was designed and synthesized by the Ruibo Biology Company (Guangdong, Guangzhou, China) as described in our previously published paper [18 (link)].

Ai K., Pan J., Zhang P., Li H., He Z., Zhang H., Li X., Li Y., Yi L., Kang Y., Wang Y., Xiang X., Chai X, & Zhang D. (2022). Methyl-CpG-binding domain protein 2 contributes to renal fibrosis through promoting polarized M1 macrophages. Cell Death & Disease, 13(2), 125.

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Exploring Cell Adhesion and ECM Remodeling

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C-phycocyanin (lot#R320190412) was purchased from Taizhou Binmei Biotechnology Co., Ltd., Taozhou, China. Recombinant human TGF-β1 (lot#0713AF354-1 L0618) was purchased from Proteintech Co., Ltd., Wuhan, China. Anti-human N-cadherin, E-cadherin, MMP-9, TGF-β ReceptorII, phospho-smad2/3, smad2/3 were purchased from Cell Signaling Technology, Inc. Danvers, MA, USA. The immunofluorescence staining kit (lot No. 13I13H10A0964) was purchased from Boster Biological Engineering Co., Ltd. All other chemicals reached analytical grade.

Ji H., Liu G., Han J., Zhu F., Dong X, & Li B. (2020). C-phycocyanin inhibits epithelial-to-mesenchymal transition in Caski cells. Cancer Cell International, 20, 292.

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Fibrotic and Inflammatory Responses of LX-2 Cells

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The immortalized human hepatic stellate cell line LX-2 was a gift from Professor Lieming Xu (Shanghai Univesity of TCM, Institute of Liver Diseases, Shuguang Hospital, Shanghai, China) 27 (link) and was detected to be free of mycoplasma. Cells were cultured in Dulbecco's modified Eagle's medium (HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum (Life Technologies, Grand Island, NY, USA) at 37°C in a humidified environment containing 5% CO2. A recombinant human TGF-β1 (Protein Tech Group, Inc, Chicago, USA) at a concentration of 10 ng/mL was added to the cell culture after FAT10 knockdown for detection of fibrotic factors and inflammatory cytokine secretion.

Zheng W., Guan F., Xu G., Yu Y., Xiao J, & Huang X. (2023). FAT10 Silencing Prevents Liver Fibrosis through Regulating SIRT1 Expression in Hepatic Stellate Cells. International Journal of Medical Sciences, 20(4), 557-565.

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TGF-β1 Induced Epithelial-Mesenchymal Transition

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HK-2, a human kidney epithelial cell line, was purchased from the Shanghai Institute for Biological Sciences (SIBS), Chinese Academy of Sciences (Shanghai, China) and cultured for less than six months from the time of resuscitation. HK-2 cells were cultivated in Dulbecco’s modified Eagle’s medium (DMEM) /F-12 1:1 (Hyclone Laboratories, Inc., Logan, UT, USA) supplemented 10% fetal bovine serum supplemented with 1% penicillin/streptomycin at 37 °C, 5% CO₂ in a humidified incubator. At ∼50% confluency, cells were treated with recombinant human TGF-β1 (10 ng/ml) (Proteintech Group, Inc.) for 24 h, 48 h, 72 h, 96 h, 120 h and 144 h or the vehicle control in DMEM/F12 1:1 for the indicated time period.

Liu G., Liu X, & Yang Y. (2022). Comparative transcriptome analysis of miRNA in hydronephrosis male children caused by ureteropelvic junction obstruction with or without renal functional injury. PeerJ, 10, e12962.

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Immortalized Human Hepatic Stellate Cell Line LX-2 Culture

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The immortalized human hepatic stellate cell line LX‐2 was obtained from Cell bank of Chinese Academy of Sciences (Shanghai, China) and cultured in Dulbecco's modified Eagle's medium (DMEM) (Sigma‐Aldrich, St. Louis, MO, United States) containing 10% fetal bovine serum (Sigma‐Aldrich) and 1% penicillin/streptomycin. The cells were incubated at 37°C in a humidified atmosphere supplied with 5% CO₂. LX‐2 cells were activated with 5 ng/mL recombinant human TGF‐β1 (Protein Tech Group, Inc, Chicago, USA). To inhibit the TGF‐β1/Smad3 signalling pathway, LX‐2 cells were treated with 10 μM SB431542 (type I TGF‐β1 receptor inhibitor) (Selleckchem, USA) for 24 h after MFAP2 overexpression.

Sun Y., Chen X., Chen L., Bao B., Li C, & Zhou Y. (2023). MFAP2 promotes HSCs activation through FBN1/TGF‐β/Smad3 pathway. Journal of Cellular and Molecular Medicine, 27(21), 3235-3246.

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