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Il 6 cytokines

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IL-6 cytokines are proteins involved in the immune response and inflammatory processes. They play a key role in the regulation of immune cells and the inflammatory cascade. The core function of IL-6 cytokines is to mediate the body's response to various stimuli, including infection and injury.

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Lab products found in correlation

Stemspan sfem 2 medium, by STEMCELL (4 mentions) Matrigel, by Corning (4 mentions) Interleukin 3 (il 3), by Thermo Fisher Scientific (4 mentions) Histopaque 1077, by Merck Group (4 mentions) Cytotune ips 2.0 sendai reprogramming vectors, by Thermo Fisher Scientific (4 mentions) Y 27632, by STEMCELL (4 mentions) Mtesr1 medium, by STEMCELL (4 mentions) Dispase, by STEMCELL (4 mentions) Cryostor cs10 freezing medium, by STEMCELL (4 mentions) Tgf β1, by Thermo Fisher Scientific (2 mentions) C6 flow cytometer, by BD (1 mentions) M csf, by Thermo Fisher Scientific (1 mentions) Intracellular staining kit, by Thermo Fisher Scientific (1 mentions) Anti human cd16, by BioLegend (1 mentions)

6 protocols using il 6 cytokines

1

Generation of induced pluripotent stem cells

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PBMCs were isolated from the peripheral blood sample using Histopaque®−1077 (Sigma-Aldrich) and cultured in StemSpan SFEMIImedium (Stem Cell Technologies) supplemented with 100 ng/mL SCF, 100 ng/mL FLT-3 L, 20 ng/mL IL-3 and 20 ng/mL IL-6 cytokines (Peprotech). Five days later, the cells were counted and transduced using CytoTune®-iPS 2.0 Sendai reprogramming vectors (Thermo Fisher) following the manufacturer’s instruction. The transduced cells were then plated onto irradiated mouse embryonic fibroblasts (MEFs) and cultured in mTeSR™1 medium (Stem Cell Technologies) which was changed every other day. Around day19 post-transduction, ESC-like colonies appeared and were manually picked on day25 post-transduction. The iPSCs were cultured on Matrigel (Corning)-coated plates in mTeSR™1 medium at 37 °C with 5% CO2 and routinely passaged at 1:3 ratio using dispase (Stem Cell Technologies) every 4–6 days. The iPSCs were frozen in CryoStor® CS10 freezing medium and thawed with 10 μM Y-27632 (Stem Cell Technologies). hESC (H7 [Wi Cell Research Institute, Madison, WI, USA]) was cultured in parallel with FDEENTi002-A.
Bai X., Yang X.J, & Chen L. (2019). Establishment of an induced pluripotent stem cell line (FDEENTi002-A) from a patient with Best’s disease carrying c.888C > A mutation in BEST1 gene. Stem cell research, 38, 101459.
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2

Generating Human iPSCs from PBMCs

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PBMCs were isolated from the whole blood sample using Histopaque®−1077 (Sigma-Aldrich) and cultured in StemSpan SFEMIImedium (Stem Cell Technologies) supplemented with 100 ng/ mL SCF, 100 ng/mL FLT-3 L, 20 ng/mL IL-3 and 20 ng/mL IL-6 cytokines (Peprotech). Five days later, the cells were collected and transduced with CytoTune®-iPS 2.0 Sendai reprogramming vectors (Thermo Fisher) following the manufacturer’s instruction. The transduced cells were plated onto irradiated mouse embryonic fibroblasts (MEFs) and maintained in mTeSR™1 medium (Stem Cell Technologies) which was changed every other day. Around day16 post-transduction, ESC-like colonies appeared and were manually picked on day20 post-transduction. The established iPSCs were cultured on Matrigel (Corning)-coated plates in mTeSR™1 medium at 37 °C with 5% CO2 and routinely passaged at 1:3 ratio using dispase (Stem Cell Technologies) every 4–6 days. The iPSCs were frozen in CryoStor® CS10 freezing medium and thawed with 10 μM Y-27632 (Stem Cell Technologies). hESC (H7 [Wi Cell Research Institute, Madison, WI, USA]) was cultured in parallel with FDEENTi003-A.
Bai X., Yang X.J, & Chen L. (2019). Generation of an integration-free induced pluripotent stem cell line (FDEENTi003-A) from a patient with pathological myopia. Stem cell research, 39, 101495.
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3

Generation of induced pluripotent stem cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
PBMCs were isolated from the peripheral blood sample using Histopaque®−1077 (Sigma-Aldrich) and cultured in StemSpan SFEMIImedium (Stem Cell Technologies) supplemented with 100 ng/mL SCF, 100 ng/mL FLT-3 L, 20 ng/mL IL-3 and 20 ng/mL IL-6 cytokines (Peprotech). Five days later, the cells were counted and transduced using CytoTune®-iPS 2.0 Sendai reprogramming vectors (Thermo Fisher) following the manufacturer’s instruction. The transduced cells were then plated onto irradiated mouse embryonic fibroblasts (MEFs) and cultured in mTeSR™1 medium (Stem Cell Technologies) which was changed every other day. Around day19 post-transduction, ESC-like colonies appeared and were manually picked on day25 post-transduction. The iPSCs were cultured on Matrigel (Corning)-coated plates in mTeSR™1 medium at 37 °C with 5% CO2 and routinely passaged at 1:3 ratio using dispase (Stem Cell Technologies) every 4–6 days. The iPSCs were frozen in CryoStor® CS10 freezing medium and thawed with 10 μM Y-27632 (Stem Cell Technologies). hESC (H7 [Wi Cell Research Institute, Madison, WI, USA]) was cultured in parallel with FDEENTi002-A.
Bai X., Yang X.J, & Chen L. (2019). Establishment of an induced pluripotent stem cell line (FDEENTi002-A) from a patient with Best’s disease carrying c.888C > A mutation in BEST1 gene. Stem cell research, 38, 101459.
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4

Cytokine Stimulation of T Cells

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TGF-β1 used was purchased from PeproTech. Working concentrations of 5ng/mL were used to stimulate T cells in all experiments. IL-6 cytokines were purchased from PeproTech and were used to stimulate T cells at a working concentration of 100ng/mL.
Lam E., Choi S.H., Pareek T.K., Kim B.G, & Letterio J.J. (2015). Cyclin-dependent kinase 5 represses Foxp3 gene expression and Treg development through specific phosphorylation of Stat3 at Serine 727. Molecular immunology, 67(2 0 0), 317-324.
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5

Generating Human iPSCs from PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
PBMCs were isolated from the whole blood sample using Histopaque®−1077 (Sigma-Aldrich) and cultured in StemSpan SFEMIImedium (Stem Cell Technologies) supplemented with 100 ng/ mL SCF, 100 ng/mL FLT-3 L, 20 ng/mL IL-3 and 20 ng/mL IL-6 cytokines (Peprotech). Five days later, the cells were collected and transduced with CytoTune®-iPS 2.0 Sendai reprogramming vectors (Thermo Fisher) following the manufacturer’s instruction. The transduced cells were plated onto irradiated mouse embryonic fibroblasts (MEFs) and maintained in mTeSR™1 medium (Stem Cell Technologies) which was changed every other day. Around day16 post-transduction, ESC-like colonies appeared and were manually picked on day20 post-transduction. The established iPSCs were cultured on Matrigel (Corning)-coated plates in mTeSR™1 medium at 37 °C with 5% CO2 and routinely passaged at 1:3 ratio using dispase (Stem Cell Technologies) every 4–6 days. The iPSCs were frozen in CryoStor® CS10 freezing medium and thawed with 10 μM Y-27632 (Stem Cell Technologies). hESC (H7 [Wi Cell Research Institute, Madison, WI, USA]) was cultured in parallel with FDEENTi003-A.
Bai X., Yang X.J, & Chen L. (2019). Generation of an integration-free induced pluripotent stem cell line (FDEENTi003-A) from a patient with pathological myopia. Stem cell research, 39, 101495.
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6

Cytokine Profiling in TBI Patients

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TGF-β1, M-CSF, and IL-6 cytokines were purchased from PeproTech. Antibodies for flow cytometry were purchased from BD (anti-human CD16), from BioLegend (anti-human LAP/latency-associated peptide TGF-β1), and from eBioscience (anti-human CD4, CD8, CD19, CD11b, and CD14, and anti-mouse CD11b, Ly6C, F4/80, CD45, CD206, CD31, and F4/80). Antibodies for the blocking experiment were purchased from BD (purified rat anti-mouse, human TGF-β1, purified NA/LE rat anti-mouse M-CSF), and from eBioscience (anti-mouse IL-6 functional grade purified). For staining surface markers, cells were suspended in PBS solution supplemented with 1% heat-inactivated fetal calf serum. For intracellular staining, cells were fixed and permeabilized with the Intracellular Staining Kit (eBioscience). Data were acquired on a C6 flow cytometer (BD) and analyzed with FlowJo software. Concentrations of M-CSF, TGF-β1, IL-6, TNF-α, G-CSF, and GM-CSF from TBI patients or mice were detected by ELISA kits from eBioscience or XITANG.
Li Z., Xiao J., Xu X., Li W., Zhong R., Qi L., Chen J., Cui G., Wang S., Zheng Y., Qiu Y., Li S., Zhou X., Lu Y., Lyu J., Zhou B., Zhou J., Jing N., Wei B., Hu J, & Wang H. (2021). M-CSF, IL-6, and TGF-β promote generation of a new subset of tissue repair macrophage for traumatic brain injury recovery. Science Advances, 7(11), eabb6260.
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