M199 and RPMI 1640 media, fetal bovine serum (FBS) and antibiotic-antimycotic mix (100X) were purchased from GIBCO/BRL (Grand Island, NY, USA); 0.25% trypsin and tryple express were acquired from INVITROGEN (Waltham, MA, USA. Sterile plastic material for tissue culture was obtained from NUNC (Rochester, NY, USA) and CORNING (Glendale, AZ, USA). Flow cytometry reagents were purchased from Becton Dickinson (Franklin Lakes, NJ, USA). Tumor necrosis factor alpha (TNF-α) was purchased from R&D Systems (Minneapolis, MN, USA). Thymidine [methyl-3H] was supplied by Perkin Elmer (Boston, MA, USA). 2,7-dichlorodihydrofluorescein diacetate (H2DCFDA) was purchased from Molecular Probes, Invitrogen (Carlsbad, CA, USA). Peroxidase-labeled monoclonal antibody against Von Willebrand factor and all the fluorescein isothiocyanate (FITC)-labeled monoclonal antibodies vs. human adhesion molecules were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). TiO2-NPs were acquired from Paris Drugstore (Mexico City, Mexico).
Tumor necrosis factor alpha tnf α
Manufactured by R&D Systems
Sourced in United States
Tumor necrosis factor alpha (TNF-α) is a cytokine produced by various cells, including macrophages, lymphocytes, and endothelial cells. It plays a crucial role in the regulation of immune responses, inflammation, and cell death.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Lipopolysaccharide lps, by Merck Group (2 mentions)
Interferon gamma ifn γ, by R&D Systems (2 mentions)
Vascular endothelial growth factor (vegf), by Merck Group (2 mentions)
Nitric oxide, by Enzo Life Sciences (2 mentions)
Malondialdehyde mda, by ZeptoMetrix (2 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions)
Flow cytometry reagent, by BD (1 mentions)
Methyl 3h thymidine, by PerkinElmer (1 mentions)
2 7 dichlorodihydrofluorescein diacetate h2dcfda, by Thermo Fisher Scientific (1 mentions)
Tryple express, by Thermo Fisher Scientific (1 mentions)
Trichloroacetic acid tca, by Merck Group (1 mentions)
4 2 68 hydroxyethyl 1 piperazineethanesulfonic acid hepes, by Thermo Fisher Scientific (1 mentions)
Trypsin 0.53 mm edta, by Thermo Fisher Scientific (1 mentions)
Dihydroethidium dhe, by Biotium (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Dithiothreitol (dtt), by Merck Group (1 mentions)
Trypan blue, by R&D Systems (1 mentions)
Interleukin il 1β, by R&D Systems (1 mentions)
14 protocols using tumor necrosis factor alpha tnf α
1
Cytotoxicity and Oxidative Stress Assay
2
Endothelial Cell Oxidative Stress Assay
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DEAE Sepharose Fast Flow, Sephadex G-75, and Coomassie brilliant blue G-250 were purchased from Solabio Technology Co. Ltd. (Beijing, China). Trichloroacetic acid (TCA), ethylenediaminetetraacetic acid disodium salt (EDTA), β-mercaptoethanol (β-M), and ammoniummum sulfate were purchased from Sigma-Aldrich (St. Louis, MO, USA). Human vascular endothelial cells of the type EA.hy926 (CRL-2922) were purchased from American Type Culture Collection (Manassas, VA, USA). Dulbecco’s modified Eagle’s medium (DMEM), nonessential amino acids (NEAA), penicillin-streptomycin (pen-strep), fetal bovine serum (FBS), 4-(2-68 hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and 0.25% (w/v) trypsin-0.53 mM EDTA were purchased from Gibco Invitrogen (Burlington, ON, Canada). Dithiothreitol (DTT) and Triton-X-100 were obtained from Sigma-Aldrich (St. Louis, MO, USA). Tumor necrosis factor-alpha (TNFα) was obtained from R&D Systems (Minneapolis, MN, USA). Dihydroethidium (DHE) was purchased from Biotium (Fremont, CA, USA).
3
Evaluating Isorhapontigenin's Anti-Inflammatory Effects
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Fetal bovine serum (FBS), penicillin-streptomycin (PS), and Dulbecco’s modified Eagle’s medium (DMEM) were purchased from Invitrogen (Carlsbad, CA, USA). Trypsin EDTA was bought from Gibco (Waltham, MA, USA). Isorhapontigenin was purchased from Sigma Chemical (St. Louis, MO, USA). Enzyme-linked immune sorbent assay (ELISA) development kits, tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and interleukin (IL-1β) were acquired from R&D Systems (Minneapolis, MN, USA). The primary antibodies α-tubulin, β-tubulin, SPHK1, SPHK2, PARP, caspase-3, caspase-9, p38, pp38, JNK, pJNK, ERK, and pERK were purchased from Cell Signaling (Beverly, MA, USA). Secondary antibodies for Sirt1, Bax, Bcl2, cytochrome-C, and GAPDH were purchased from Santa Cruz Technology. MCF7, T47D, and MDA-MB-231 cells were purchased from the Korean Cell Line Bank. 3-[4,5-Dimethyl-2-thiazolyl]-2,5-diphenyl-2-tetrazolium bromide (MTT) powder, RNase-A, propidium iodide, and DCFDA were purchased from Sigma-Aldrich (St. Louis, MO, USA). The annexin V-FITC apoptosis detection kit and trypan blue were purchased from R and D Systems.
4
Glyceryl Tributyrate Dietary Modulation
5
Cell Proliferation and Differentiation
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AP was bought from Sigma-Aldrich (USA), and Alpha minimum Eagle's medium (Alpha-MEM), foetal bovine serum and penicillin were bought from Gibco BRL (Gaithersburg, MD, USA). However, the CCK-8 assay kit was bought from Dojindo Molecular Technology (Japan). Tumor necrosis factor alpha (TNF-α) and BMP-2 were bought from R&D Systems (USA).
6
Investigating Cell Signaling Pathways
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MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide), antibody to β-actin, and lipopolysaccharide (LPS) were purchased from Sigma (St Louis, MO, USA). The antibodies to phosphorylated ERK (pERK1/2), ERK1/2, pJNK, JNK, pFAK, and FAK were purchased from Cell Signaling Technology (Beverly, MA, USA), antibodies to RhoA, Cdc42, and GAPDH were purchased from Santa Cruz (Santa Cruz, CA, USA), and antibody to MMP-9 was obtained from Millipore (Billerica, MA, USA). Platelet-derived growth factor (PDGF-BB) and tumor necrosis factor alpha (TNF-α) were obtained from R & D systems (Minneapolis, USA). Nitrocellulose membrane and chemiluminescent for Western blotting were purchased from Bio-Rad (Richmond, CA, USA) and IMEGENEX (San Diego, CA, USA), respectively. All solvents, chemicals, and reagents were of analytical grade and purchased from Sigma-Aldrich unless otherwise specified.
7
Apigenin and Methylcellulose Modulate Inflammatory Pathways
8
Autologous Dendritic Cell Vaccine Production
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AGS-003 was manufactured at a centralized GMP compliant facility (Argos Therapeutics, Durham, NC). Following screening and consent, autologous tumor total RNA was isolated from nephrectomy or metastasectomy tissue samples and messenger RNA was amplified using RT/PCR and in vitro transcription technologies as previously described [47 (link)]. CD40L RNA was manufactured using in vitro transcription and a post-transcriptional capping method [48 (link)]. Patients had leukapheresis at the clinical site’s donor center using a COBE Spectra® Leukapheresis System (Gambro BCT, Lakewood, CO). Monocytes were cultured in AIM-V media with 800 U/mL granulocyte macrophage-colony stimulating factor (Berlex) and 1000 U/mL IL-4 (R&D Systems) to generate immature DCs that were then matured using 20 ng/mL tumor necrosis factor alpha (TNF α) (R&D Systems)/1000 U/mL IFN-γ (InterMune)/1 μg/mL prostaglandin E2 (Sigma). Mature DCs were electroporated with the amplified tumor RNA and CD40L RNA using a post-maturation electroporation protocol [10 (link)].
The final AGS-003 product was formulated as 1.4 × 107 DC/0.7 mL in 80% autologous plasma, 10% dextrose (50% w/v) (Hospira), and 10% DMSO (Sigma) and cryopreserved in liquid nitrogen vapor phase. Thawed samples of final product were assessed for sterility, mycoplasma, endotoxin, and viability prior to release for clinical use.
The final AGS-003 product was formulated as 1.4 × 107 DC/0.7 mL in 80% autologous plasma, 10% dextrose (50% w/v) (Hospira), and 10% DMSO (Sigma) and cryopreserved in liquid nitrogen vapor phase. Thawed samples of final product were assessed for sterility, mycoplasma, endotoxin, and viability prior to release for clinical use.
9
HA-based Biomaterial Characterization
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HA (90 kDa) was kindly donated by JNC Corporation (Japan). EGCG was obtained from DSM Nutritional Products (Switzerland). Dextran (100 kDa), bovine serum albumin (BSA), phosphate-buffered saline (PBS) and incomplete Freund's adjuvant were purchased from Sigma-Aldrich (USA). Fetal bovine serum (FBS) and PicoGreen dsDNA assay kit were purchased from Thermo Fisher (Singapore). Tumor necrosis factor-alpha (TNFα) was purchased from R&D System (USA). Bovine type II collagen was obtained from Chondrex (USA). 2-(N-Morpholino)ethanesulfonic acid (MES), N-hydroxysuccinimide (NHS), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl), 5-aminofluorescein (AF) and 4,6-diamidino-2-phenylindole (DAPI) were purchased from Sigma-Aldrich (Singapore). 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMTMM) was acquired from Tokyo Chemical Industry Co. Ltd. (Japan). Cyanine7.5 amine dye (Cy7.5) was obtained from Lumiprobe Corporation (USA). Rat anti-human CD44 antibody (clone Hermes-1), isotype control antibody, fluorescein isothiocyanate (FITC)-labeled secondary antibody, Pierce™ quantitative peroxide assay kit and alamarBlue® cell viability assay reagent were purchased from Thermo Fisher Scientific (Singapore). CellTiter-Glo® luminescent cell viability assay reagent was obtained from Promega (Singapore).
10
Generating Antigen-Specific CD8+ T Cells from Peripheral Blood
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CD8+ or CD14+ cells were enriched by human CD8 or CD14 MicroBeads (Miltenyi Biotec), respectively. CD14+ cells were cultured with 100 ng/mL GM-CSF (R&D Systems) and 25 ng/mL interleukin-4 (IL-4) (R&D systems) for 6 days to differentiate into DCs. On day 5, 10 ng/mL IL-6 (R&D Systems), 10 ng/mL IL-1β (R&D Systems), 10 ng/mL tumor necrosis factor alpha (TNF-α) (R&D Systems), and 1 μg/mL PGE2 (Sigma-Aldrich, P5640, Tokyo, Japan) were added for DC maturation. All cells were cryopreserved unless they were required for immediate use. For the first round of stimulation, 5 × 105 CD8+ cells were cocultured with 5 × 104 DCs in the presence of pooled peptides (3.3 μg/mL each peptide) for 12 days. 100 U/mL IL-2 (Shionogi), 5 ng/mL IL-7 (PeproTech, Cranbury, NJ, USA), and 5 ng/mL IL-15 (PeproTech) were added from day 3. For the second round, proliferated cells were restimulated with a 10–50:1 ratio of DCs. Cytokine-containing fresh medium was periodically added every 2 or 3 days. For the third or fourth round, peptide reactive cells were further cocultured with a 5–50:1 ratio of DCs with a 3 μg/mL single peptide. Responses to corresponding single peptides were monitored by IFN-γ ELISPOT assay or CD137 upregulation by fluorescence-activated cell sorting (FACS) 20–24 h after C1R-A∗24:02 and 10 μg/mL peptide stimulation.
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