Fibronectin and octadecyl rhodamine B chloride (R18) were obtained from Sigma-Aldrich (St. Louis, MO, United States) and Invitrogen (Carlsbad, CA, United States), respectively. IL-1β, IL-6, and TNF-α were obtained from Sino Biological (Beijing, China). Adhesive substrates were prepared from perfusion of microfluidic channels in the BioFlux plate with different concentration of FN at 5 dyn/cm2 for 15 min, followed by incubation at RT for 1 h, and blocked with 0.5% BSA at 5 dyn/cm2 for 15 min. All cytokine-containing adhesive substrates were prepared from perfusion of cytokines together with FN. MSCs were trypsinized, stained with 300 nM R18, resuspended at 1 × 106 cells/ml in PBS, and perfused from the inlet wells for 10 min at 10 dyn/cm2. In experiments that assess the effect of soluble cytokines, cells were incubated with different cytokines for 30 min at RT before perfusion. Images of attached cells after the perfusion were captured at a magnification of 100× using a Nikon TS100 microscope (Nikon Instruments, Inc., Melville, NY, United States) equipped with a CCD camera (QICAM, QImaging, Surrey, British Columbia) and the BioFlux 200 software. Cell number from each image was counted and averaged for each group.
Interleukin 6 (il 6)
Manufactured by Sino Biological
Sourced in China, United States
IL-6 is a recombinant protein produced by Sino Biological. It is a member of the interleukin cytokine family and plays a central role in the immune response and inflammation.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 3 (il 3), by Sino Biological (3 mentions)
Il 1β, by Sino Biological (2 mentions)
Flt3l, by Thermo Fisher Scientific (2 mentions)
Stem cell factor (scf), by Thermo Fisher Scientific (2 mentions)
Bone morphogenetic protein 4 (bmp4), by Thermo Fisher Scientific (2 mentions)
Basic fibroblast growth factor (bfgf), by Sino Biological (2 mentions)
Chir99021, by Selleck Chemicals (2 mentions)
Activin a, by Sino Biological (2 mentions)
Ly294002, by Selleck Chemicals (2 mentions)
Tnf α, by R&D Systems (2 mentions)
Fibronectin, by Merck Group (1 mentions)
Ts100 microscope, by Nikon (1 mentions)
Tnf α, by Sino Biological (1 mentions)
Octadecyl rhodamine b chloride r18, by Merck Group (1 mentions)
A83 01, by Selleck Chemicals (1 mentions)
Growth factor reduced matrigel, by BD (1 mentions)
Iwr 1 endo, by Selleck Chemicals (1 mentions)
Vascular endothelial growth factor (vegf), by Selleck Chemicals (1 mentions)
Basic fibroblast growth factor (bfgf), by Selleck Chemicals (1 mentions)
Sb431542, by Selleck Chemicals (1 mentions)
14 protocols using interleukin 6 (il 6)
1
Adhesion of MSCs on Cytokine-Coated Substrates
2
Hematopoietic Differentiation of H1 hESCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hematopoietic differentiation was performed as previously described (Zhu et al., 2020 (link)). Briefly, H1 hESCs were plated onto Growth Factor Reduced Matrigel (1:200 dilution; BD Biosciences)-coated plates at a proper initial density. The hematopoietic differentiation medium was changed every day, supplemented with corresponding cytokines or inhibitors, and floated hESCs-HSPCs were collected from D8 for further assays. The cytokines or inhibitors added each day were as follows: D0–D1, 40 ng/mL BMP4 (PeproTech), 30 ng/mL Activin A (Sino Biological), 20 ng/mL bFGF (Sino Biological), 6 μM CHIR99021 (Selleck), and 10 μM LY294002 (Selleck); D1–D2, 30 ng/mL BMP4, 1 μM A8301 (Selleck), and 2 μM IWR-1-endo (Selleck); D2–D4, 40 ng/mL vascular endothelial growth factor (VEGF) (Sino Biological) and 50 ng/mL bFGF; D4 and later, 40 ng/mL VEGF, 50 ng/mL bFGF, 10 μM SB431542 (Selleck), 10 ng/mL SCF (PeproTech), 50 ng/mL TPO (Sino Biological), 10 ng/mL IL3 (Sino Biological), 50 ng/mL IL6 (Sino Biological), and 50 ng/mL Flt3L (PeproTech).
3
IL-6 and TNF-Induced Adipocyte Migration Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mature adipocytes (4 × 104 cells) differentiated from 3T3-L1 preadipocytes were seeded in 12-well culture plates and treated with IL-6 (20 ng/mL) and TNF (10 ng/mL) (Sino Biological, Beijing, China) for 24 h. After washing twice with PBS, the cells were cultured in DMEM supplemented with 2% FBS in the presence of 100 μM 3-OH phloretin or phloretin, as described previously [19 (link)]. The purified bone marrow-derived M1 macrophages (1 × 105 cells), isolated using magnetic-activated cell sorting (MACS), were added to the upper chamber of a transwell plate (3 μM; Corning Life Sciences, Corning, NY, USA) and incubated for 48 h. The migrated cells were washed with PBS before fixation with 4% paraformaldehyde. The cells stained with hematoxylin were counted.
4
Cell Line Cultivation and Drug Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HEK293T and U2OS cell lines were grown in the DMEM containing 10% FBS and 2 mM L-glutamine (Biosera, Kansas City, MO, USA). The HEK293 cell line was grown in the IMDM containing 10% FBS and 2 mM L-glutamine. Human MM lines OPM-2 and RPMI8226 purchased from the Leibniz Institute DSMZ (Germany) were grown in RPMI 1640 (Biosera) supplemented with 10% FBS, 2 mM L-glutamine, and 2 ng/mL IL-6 (Sino Biological, Wayne, PA, USA). All cell lines used in the study were tested for mycoplasma contamination. Cells were treated with bortezomib (BTZ, UBPBio, PS341), MG132 (Sigma, M7449), or carfilzomib (CFZ, UBPBio, PR171) at various concentrations as indicated. Lopinavir (LPV), darunavir (DRV), brecanavir (BCV), tipranavir (TPV), amprenavir (APV), nelfinavir (NFV), indinavir (IDV), and ritonavir (RTV) were kindly provided by Dr. J. Konvalinka (Prague, Czech Republic).
5
Quantitative Detection of Inflammatory Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the detection of serum AEBP1- (CLOUDCLONE CORP, Wuhan, China) and VSMC-secreted AEBP1, TNF-α (R&D Systems, Minneapolis, MN), IL-10 (R&D Systems, Minneapolis, MN), IL-6 (Sino Biological, Beijing, China), and IL-1β (Sino Biological, Beijing, China) in the cell culture after 24 h, we used commercial enzyme-linked immunosorbent assay (ELISA) kits following the manufacturer's guidelines. Briefly, 100 µL of the sample was used per well, and the plate was incubated with the antibodies for 1 h at room temperature. After the incubation, the wells were washed three times, and 100 µL of each chromogenic substrates A and B was added. The plate was then incubated in the dark for 20 min at 37°C. Finally, 50 µL of a stop solution was added to each well to stop the reaction, and the absorbance was measured at 450 nm. Expression values were calculated using a standard curve.
6
Bone Marrow-Derived Macrophages and Myeloid-Derived Suppressor Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow cells were obtained by flushing the femoral bone marrow cavity of C57BL/6 mice with RPMI 1640 (Gbico, C11875500BT) medium. Red blood cells were removed using red blood cell lysis buffer (Beyotime, C3702) and washed once with PBS. For generation of bone marrow‐derived macrophages (BMDMs), the bone marrow cells were then cultured in RPMI 1640 containing 10% FBS, 10 ng mL−1 M‐CSF (Sino biological, 51112‐MNAH) for 7 days to obtain bone marrow‐derived macrophages. For generation of bone marrow myeloid‐derived suppressor cells (BM‐MDSCs), the bone marrow cells were then cultured in RPMI 1640 containing 10% FBS, 20 ng mL−1 GM‐CSF (Sino biological, 51048‐MNAH) and 20 ng mL−1 IL‐6 (Sino biological, 50136‐MNAE) for 5 days to obtain bone marrow myeloid‐derived suppressor cells.
7
Cytokine and TLR7/8 signaling in BMDMs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
rIL6: BMDM were treated with IL-6 (Cat#50136-MNAE, Sino Biological) for along with 1 µg/mL R848 for 4 h. IFN-I: BMDMs were pre-conditioned with IFN-alpha4 (Cat#50672-M08H, Sino Biological) for 16, 24, or 48 h, and then activated with R848(1 µg/ml) for 4 h. The supernatant was harvested and stored in −80 °C refrigerator for subsequent ELISA measurement. Anti-IL-6: 3.5 × 105 sorted (purity>99%) Flt3L-pDC activated with 1 µg/ml R848 containing in 1.4 ml RP10 for 24 h. Then Flt3L-pDC supernatants were pre-incubated with 1 µg/ml anti-IL6 mIgG (Cat#mabg-mIL6-3, Invivogen) for 30 min. 3 × 105 BMDMs were co-cultured with 50% Flt3l-pDC supernatant or anti-IL6 pre-treated Flt3L-pDC supernatant in the presence of 1 µg/ml R848 and 10 µg/ml brefeldin A for 4 h, and were analyzed by intracellular staining with FACS.
8
Modulating IL-6 Signaling in TF-1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TF-1 cells were seeded in flat-bottomed, 96-well plates at 10,000 cells/well in the presence of 1, 10, or 50 ng/mL IL-6 (Sino Biological, Beijing, China). The cells were then incubated with 0.1, 1, or 10 μg/mL of scFv-IL6R-Fc or tocilizumab (Roche) for 48 h. Following incubation, cell proliferation was measured by the Cell Counting Kit (CCK)-8 assay (Dojindo Molecular Technologies, Shanghai, China) according to the manufacturer’s protocol.
9
Stepwise Differentiation of hPSCs to Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hPSCs were dissociated by Accutase (Sigma) and plated on growth factor-reduced Matrigel (Corning)-coated plates with thiazovivin (0.1 μM, Selleck). hPSCs were induced for stepwise blood differentiation in basal medium supplemented with cytokines and inhibitors.23 (link) Briefly, hPSCs were treated with DMEM/F12 (Gibco) medium containing 40 ng/mL BMP4 (Peprotech), 30 ng/mL ACTIVIN A (Sino Biological), 20 ng/mL basic fibroblast growth factor (bFGF) (Sino Biological), 6 μM CHIR99021 (Selleck), and 10 μM LY294002 (Selleck) for 1–2 days. Then, we switched to medium containing 40 ng/mL vascular endothelial growth factor (Sino Biological) and 50 ng/mL bFGF for 2 or 3 days. HSPCs were induced by medium containing 10 ng/mL SCF (Peprotech), 50 ng/mL thrombopoietin (Sino Biological), 10 ng/mL IL-3 (Sino Biological), 50 ng/mL IL-6 (Sino Biological), and 50 ng/mL FMS-like tyrosine kinase 3 ligand (FLT3L) (Peprotech). Floating iHSPCs were collected and plated in myeloid differentiation medium (StemPro +50 ng/mL FLT3L + 50 ng/mL M-CSF + 25 ng/mL GM-CSF) for 12 days and switched to macrophage maturation medium (RPMI-1640 + 10% FBS + 50 ng/mL M-CSF) for 7 days. All of the recombinant human cytokines were purchased from Sino Biological.
10
Heparin and Heparan Sulfate Modulation of Cytokine Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cytokines used in this study include IL‐2 (PeproTech, USA), IL‐1beta, IL‐6 and CCL8 (Sino Biological, China or PeproTach, USA). Heparin (MW≈12 kDa) was obtained from SPH No.1 Biochemical Pharmaceutical CO., LTD, China. OD‐heparin (O‐desulfated heparin; MW≈12 kDa), NS‐K5 (N‐sulfated K5 polysaccharide; MW≈60 kDa) were prepared as described,[24] (link) HS (heparan sulfate; MW≈30 kDa) was purified from bovine lung.[25] (link)
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!