Naïve CD4+ CD62L+ T cells from the spleens of unprimed OT-II TCR transgenic mice were purified using the MACS cell sorting system (Miltenyi Biotec). CD4+ CD62L+ T cells (7.0×105) were stimulated with 5 µg/mL plate-bound anti-CD3 Abs (BD Pharmingen) and 1 µg/ml soluble anti-CD28 Abs (BD Pharmingen) for three days in RPMI-1640 supplemented with 10% fetal bovine serum and 2-ME in the presence of recombinant cytokines. T cells were polarized with 10 ng/mL recombinant mouse IL-12 (R&D Systems), 5 ng/ml mouse IFN-γ (R&D Systems), 20 U/mL human IL-2, and 5 µg/mL anti-IL-4 (BD Pharmingen) for Th1 cell differentiation, 5 ng/mL human TGF-β1 (PeproTech), 20 ng/mL mouse IL-6 (R&D Systems), 20 U/mL human IL-2, and anti-IL-4 and anti- IFN-γ (2 µg/mL; BD Pharmingen) for Th17 cell differentiation, as well as 5.0 ng/mL human TGF-β1 (PeproTech), 100 U/mL human IL-2, anti-IL-4, and anti- IFN-γ (5 µg/mL; BD Pharmingen) for induced Treg (iTreg) cell differentiation.
Human tgf β1
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom, China, Germany
Human TGF-β1 is a recombinant protein that represents the mature, biologically active form of the human Transforming Growth Factor beta 1 (TGF-β1) cytokine. TGF-β1 is a multifunctional cytokine that regulates a variety of cellular processes, including cell growth, differentiation, and immune function.
Automatically generated - may contain errors
Lab products found in correlation
Mouse il 6, by Thermo Fisher Scientific (6 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (5 mentions)
Il 12, by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Lipopolysaccharides (lps), by Merck Group (3 mentions)
Interleukin 2 (il 2), by Thermo Fisher Scientific (2 mentions)
Hamster igg, by MP Biomedicals (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Mouse il 6, by BD (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Il 10, by Thermo Fisher Scientific (2 mentions)
Tgf β1, by Thermo Fisher Scientific (2 mentions)
Ifn γ, by Thermo Fisher Scientific (2 mentions)
Dexamethasone (dex), by Merck Group (2 mentions)
Complete rpmi, by Thermo Fisher Scientific (2 mentions)
Mil 2, by Thermo Fisher Scientific (2 mentions)
Anti mcd3, by BioXCell (2 mentions)
Anti ifn γ, by Thermo Fisher Scientific (2 mentions)
Anti il 4, by Thermo Fisher Scientific (2 mentions)
48 protocols using human tgf β1
1
Differential T Cell Polarization
2
Murine Naive CD4+ T Cell Differentiation
3
Naive CD4+ T Cell Polarization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Naïve CD4+ (CD25-CD4+ CD25-CD62L+ CD44-) T cells were isolated from the spleens and the lymph nodes of mice by FACS sorting. 96-well T-bottom plates were pre-coated with 50 μL of hamster IgG (MP Biomedicals) at 37 °C for 1 hour. Following multiple washes with 1XDPBS, 40,000 naive CD4+ T cells were seeded in T cell media (RPMI supplemented with 10% fetal bovine serum, 25 mM glutamine, 55 µM 2-mercaptoethanol, 100 U/mL penicillin, 100 mg/mL streptomycin) and their T cell receptor downstream signaling pathways (TCR activation) were activated with soluble anti-CD3 (clone 145-2C11, 0.25 µg/mL) and anti-CD28 (clone 37.51, 1 µg/mL). For T H 1 cell differentiation, 100 U/mL of IL-2 (Peprotech) and 10 ng/mL of IL-12 (Peprotech) were added. For T H 17 cell differentiation, IL-6 (eBioscience, 20 ng/mL) and human TGF-β1 (Peprotech, 0.3 ng/mL) were added. For T reg culture, 100 U/mL of IL-2 (Peprotech) and human TGF-β1 (Peprotech, 5 ng/mL) were added. Bacterial culture supernatants or small molecules including bile acids were added 18 hours after TCR activation. Cells were harvested and assayed by flow cytometry on day 3.
4
Cell Culture Conditions for Fibroblasts, Endothelial, and Monocytic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human dermal fibroblast cell line (BJ), human dermal microvascular endothelial cell line (HMEC-1) [31 (link)], and human acute monocytic leukemia cell line (THP-1) were purchased from ATCC (Manassas, VA, USA). BJ cells were cultured in DMEM (Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS) (Gibco, Grand Island, NY, USA), 100 U/ml penicillin and 100 μg/ml streptomycin (Corning Incorporated, Corning, NY, USA) at 37 °C in a humidified 5% CO2 atmosphere. After adhering the cells, they were starved for 24 h with 0.1% FBS in DMEM, then stimulated with human TGF-β1 (PeproTech, Rocky Hill, NJ, USA) for 24 h. HMEC-1 cells were cultured in MCDB131 (without L-glutamine) as base medium (Sigma-Aldrich, St. Louis, MO, USA), supplemented with 10 ng/ml epidermal growth factor (Sigma-Aldrich, St. Louis, MO, USA), 1 μg/ml hydrocortisone (Sigma-Aldrich, St. Louis, MO, USA), 10 mM glutamine (Sigma-Aldrich, St. Louis, MO, USA),100 U/ml penicillin, 100 μg/ml streptomycin, and 10% FBS in 70 cm2 flasks until confluence at 37 °C, 5% CO2 atmosphere. After the cells were attached, TNF-α is stimulated for 24 h. THP-1 cells were cultured in RPMI 1640 (Gibco, Grand Island, NY, USA) containing 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin at 37 °C in a humidified 5% CO2 atmosphere.
5
Naive CD4+ T Cell Differentiation
6
Quantifying Tumor Spheroid Growth and Necrosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded at 1000 cells/well in round‐bottomed ultra‐low attachment (ULA) 96‐well‐plates (VWR, #4441020) in 200 μL of the appropriate medium and grown in a CO2‐incubator at 37°C. Medium was changed every 2‐3 days. Images were acquired on the indicated days with an Olympus IX83 microscope, using a SP 10× objective and CellSens Dimension software. Spheroid area was determined in ImageJ software using freehand selection to mark spheroid outline, followed by area quantification in ImageJ. Necrotic core quantification (in three independent biological replicates) was performed using ImageJ software. Bright field images were analyzed using a macros function where images were converted to 8‐bit format, adjusted to Threshold (0.50) to define the necrotic core in each spheroid. Area in pixels2 was measured using “Analyze Particles” function with size of particles bigger than 2000 pixels2, and converted to mm2. In each experiment 4‐6 biological replicates were used. Where indicated, media were supplemented with SB 431542 (#1614, Tocris Bioscience), human TGFβ1 (#100‐21‐2UG, PeproTech), Capivasertib (AZD5363, #S8019, Selleckchem), or MK‐2206 (#S1078, Selleckchem).
7
TGF-β1 and Sorafenib Effects on Ovarian Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human ovarian cancer SKOV-3 cells were obtained from the Shanghai Institute of Cell Biology (China) and maintained in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS; Gibco) in a CO2 incubator at 37°C. The cells were passed once every 2–4 days.
There were 5 cell treatment groups (n = 3 per cell treatment group), i.e., SKOV-3 cell control, TGF-β1 (10 ng/mL)-alone treatment according to previous studies [28 (link),29 ,30 (link)], sorafenib (10 µM)-alone, sorafenib (10 µM) + TGF-β1 (10 ng/mL), and TGF-β1 (10 ng/mL) + Ly2157299 (a selective TGF-β1 inhibitor, 5 µM) group. Human TGF-β1 was obtained from Pepro Tech (Shuzhou, China), while sorafenib and Ly2157299 were from TargetMol (Shanghai, China).
There were 5 cell treatment groups (n = 3 per cell treatment group), i.e., SKOV-3 cell control, TGF-β1 (10 ng/mL)-alone treatment according to previous studies [28 (link),29 ,30 (link)], sorafenib (10 µM)-alone, sorafenib (10 µM) + TGF-β1 (10 ng/mL), and TGF-β1 (10 ng/mL) + Ly2157299 (a selective TGF-β1 inhibitor, 5 µM) group. Human TGF-β1 was obtained from Pepro Tech (Shuzhou, China), while sorafenib and Ly2157299 were from TargetMol (Shanghai, China).
8
Primary Adult Microglia Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary adult microglia culture was generated as previously described (Butovsky et al., 2014 (link)). Briefly, purified adult microglia were cultured in the presence of 15% LSM media (Otero et al., 2009 (link)) and 10ng/mL human TGF-β1 (PeproTech) for 7 days before experiments. For details on in vitro assays performed, see the Extended Experimental Procedures.
9
Modulation of HSF Proliferation by PD and TGF-β1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HSFs were seeded in 6-well plates at a concentration of 1 × 105/mL in 2 mL of complete medium (CM) per well. A total of 15 wells were divided into five groups (n = 3), and each group was treated with 0, 2, 4, 6 or 8 μM PD. A total of 18 wells were divided into six groups (n = 3), and each group was treated with PD (0 μM), TGF-β1 (5 ng/mL), TGF-β1 (5 ng/mL) + PD (2 μM), TGF-β1 (5 ng/mL) + PD (4 μΜ), TGF-β1 (5 ng/mL) + PD (6 μM) or TGF-β1 (5 ng/mL) + PD (8 μΜ). PD was purchased from Yuanye Bio-Technology (Shanghai, China) and dissolved in ddH2O to prepare a 10 mM stock solution. Human TGF-β1 (Peprotech, USA) was diluted to prepare a 20 ng/mL stock solution.
10
Inhibiting 11β-HSD1 Ameliorates TGF-β1-Induced Fibrosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 11β-HSD1 inhibitor, BVT.2733, was purchased from MedChem Express. Rat TGF-β1 was purchased from Abbkine Scientific Co., Ltd. (cat. no. PRP100618). The anti-11β-HSD1 antibody was purchased from AtaGenix (cat. no. ATA24000; 1:200). Human TGF-β1 was purchased from PeproTech, Inc. (cat. no. AF-100-21). Rat Col IV (cat. no. MBS704982) and PIIINP ELISA kits (cat. no. MBS704287), and the Annexin V-FITC/PI apoptosis detection kit (cat. no. MBS355226) were purchased from MyBioSource, Inc. The HA ELISA kit was purchased from R&D Systems (cat. no. DHYAL0). The ProCell collagen colorimetric assay kit was purchased from Genmed Scientifics, Inc. (cat. no. GMS10373.8). The rabbit-derived rat α-SMA (cat. no. ab124964; 1:1,500), CTGF (cat. no. ab6992; 1:2,000) and β-actin (cat. no. ab8227; 1:1,000) primary antibodies, and the goat anti-rabbit secondary IgG-HRP antibody (cat. no. ab97080; 1:5,000) were all purchased from Abcam. The cortisol kit (11β-HSD1 activity detection kit) was purchased from Cisbio (cat. no. 62CRTPEG). The adenovirus and adenovirus vector were purchased from Miao Ling Biotechnology Co., Ltd. (cat. nos. P0241 and P0653). The Cell Counting Kit-8 (CCK-8) cell proliferation test kit (cat. no. CK04) was purchased from Dojindo Molecular Technologies, Inc.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!