Treatment was initiated after the onset of disease (on day 28 after the first immunization), when arthritis had become established (arthritis score ≥ 2). All the treatments were administered intraperitoneally. CIA mice were randomly assigned to four groups (n = 8/group). In the UC-MSC transplantation group, UC-MSCs of 5 × 106 cells were injected intraperitoneally twice, on day 28 and day 56 after the first immunization, respectively. In the anti-TNF-α-treated group, anti-TNF-α (eBioscience, USA) was administrated at a total dose of 200 μg/mouse, for 50 μg/mouse every two weeks. The zoledronic acid (ZA, Chiatai Tianqing, China) group was treated with 150 μg/kg ZA for once. The control group was treated with PBS. All animals were sacrificed after initial treatment for 8 weeks.
Anti tnf α
Manufactured by Thermo Fisher Scientific
Sourced in United States
Anti-TNF-α is a laboratory product used to detect and measure the presence of tumor necrosis factor-alpha (TNF-α) in biological samples. TNF-α is a cytokine that plays a role in inflammation and immune response. This product is designed to facilitate the quantitative analysis of TNF-α levels, which can be useful for research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti ifn γ, by Thermo Fisher Scientific (11 mentions)
Anti il 6, by Thermo Fisher Scientific (10 mentions)
Anti il 1β, by Thermo Fisher Scientific (5 mentions)
Anti il 10, by Thermo Fisher Scientific (4 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (4 mentions)
Cytofix cytoperm kit, by BD (3 mentions)
Saponin, by Merck Group (2 mentions)
Tnf α, by R&D Systems (2 mentions)
Ifn γ, by R&D Systems (2 mentions)
Anti cd62l, by Thermo Fisher Scientific (2 mentions)
Anti granzyme b, by Thermo Fisher Scientific (2 mentions)
Anti cd4, by BD (2 mentions)
Anti cd86, by BD (2 mentions)
Anti il 2, by BD (2 mentions)
Golgiplug, by BD (2 mentions)
Anti ifn γ, by BD (2 mentions)
Anti granzyme b, by BioLegend (2 mentions)
Anti cd80, by Thermo Fisher Scientific (2 mentions)
Pe conjugated anti ifn γ, by BD (2 mentions)
Anti foxp3, by Thermo Fisher Scientific (2 mentions)
42 protocols using anti tnf α
1
Mesenchymal Stem Cell Therapy for Arthritis
2
SARS-CoV-2 Peptide Stimulation of PBMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMCs were isolated from heparinized blood samples and stimulated with SARS-CoV-2 peptide pool or medium in the presence of 2 μM monensin (eBioscience, San Diego, Calif) for 24 hours. After culture, the cells were collected for flow cytometry analysis. Fluorescence-labeled mAbs against the following antigens were added to the cell suspensions as follows: fixable viability stain, CD3, CD4, CD45RA, CCR7, HLA-DR, PD-1, Tim-3, TIGIT, CTLA-4, and CD39 (BD Biosciences, San Jose, Calif). All these cell suspensions were incubated for 30 minutes on ice. In some experiments, cells were fixed and permeabilized, and stained with anti–IL-2, anti–IFN-γ, anti–TNF-α, anti–IL-4, and anti–IL-17 mAbs (eBioscience). Isotype controls with irrelevant specificities were included as negative controls. fixable viability stain was used to exclude dead cells from analysis. After washings, the pellets were resuspended in 300 μL staining buffer, followed by analysis with FACSCanto flow cytometer (BD Biosciences).
3
Quantifying SARS-CoV-2 Specific T Cell Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As described previously [28 (link)], splenocytes were incubated with SARS-CoV-2 S peptide pools (1 μg/mL, Miltenyi Biotec, Auburn, CA, USA) for 5 h in the presence of BD GolgiPlug (BD Biosciences, San Jose, CA, USA). Briefly, cells were stained with antibodies for CD4 or CD8, fixed in 2% paraformaldehyde, and permeabilized with 0.5% saponin before adding anti-IFN-γ, anti-TNF-α, or control rat IgG1 (e-Biosciences). Samples were processed with a C6 Flow Cytometer instrument. Dead cells were excluded based on forward and side light scatter. Data were analyzed with a CFlow Plus Flow Cytometer (BD Biosciences, San Jose, CA, USA).
4
Phenotyping and Functional Analysis of NK Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NK cell lines from the 3 characterized healthy donors were incubated with peptide-pulse HLA-transfected 721.221-ICP47 cells at an effector:target (E:T) cell ratio of 1:1, in the presence of anti-CD107a mAb (#H4A3; Becton Dickinson) to measure degranulation. Cells were thereafter incubated for 5 h in the presence of Golgi Stop and Golgi Plug solutions (BD Biosciences) and then stained with cell-surface markers (anti-CD3, anti-CD56, and anti-CD45). Cells were fixed, permeabilized with a cytofix/cytoperm kit (Becton Dickinson), and then intracellularly stained with anti-IFN-γ (#B27; Becton Dickinson) and anti-TNF-α (#Mab11; eBiosciences), as previously described [31 ]. Data were analyzed with the “Boolean gate” algorithm using Flow Jo version 9 (TreeStar). Pestle software was used to remove the background, and pie charts, generated with Spice software (NIAI freeware) [32 (link)], present the frequency of NK cells positive for 0, 1, 2, or 3 responses. Arcs depict the frequency of cells positive for CD107a, IFN-γ, or TNF-α.
5
Cytokine-induced Inflammatory Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human SFMSCs were seeded into 10-mm plates and cultured until they reached 80% confluence. The cells were treated with cytokines and/or inhibitors. Inhibitors [BAY11-7082 (Cat. no. 19542-67-7; Santa Cruz Biotechnology, Inc.), anti-TNF-α (167348; Ebioscience, San Diego, CA, USA)] were added 1 h prior to cytokine treatment. The empty plate using the same culture medium, but without cells as the negative control. IL-6 and IL-8 expression in the culture medium was determined using a CBA kit (Human Inflammatory Cytokine kit; Cat. no. 551811; BD Biosciences). Double-stranded DNA was detected using Quant-iT PicoGreen dsDNA reagent (Invitrogen, Carlsbad, CA, USA) as an endogenous control.
6
T Cell Response to Influenza Virus in RIG-I Knockout Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow derived dendritic cells (BMDC) were generated from RIG-I+/+ and RIG-I-/- mice and T cell re-stimulation experiments were performed as previously described[31 (link)], [32 (link)]. Briefly, BMDC were first infected with PR8 at an MOI of 0.5 for 5h, washed with PBS 3 times to remove free virus, and resuspended in Iscove's Modified Dulbecco's Media complete media with FBS (IMDM, Invitrogen). For some experiments, infected BMDC were treated with IFN-I (200U/ml) before T cells were added. T cells from naïve or IAV infected RIG-I+/+ and RIG-I-/- mice (day 7 post infection) were enriched using Pan T cells isolation kit II (Miltenyi Biotec) and co-cultured with PR8 infected BMDC at a ratio of 10:1 for 2–3 hours followed by the addition of Brefeldin A (5μg/ml; eBiosciences). DC-T cell co-cultures were further incubated for an additional 8-10hrs. The cells were first surface stained for cell surface markers, followed by intracellular staining for cytokines. For intracellular staining, cells were incubated in permeabilization and fixation buffer (BD Pharmingen) for 45 minutes followed by 2 washes in 1xPBS, 1%FBS, 0.5% Saponin (Sigma, St Louis, MO) and intracellular staining with anti-IFNγ (2μg/ml, XMG1.2) (Biolegend), anti- Granzyme B (2μg/ml, GB11) (Biolegend), anti-TNFα (2μg/ml, MP6-XT22) (eBiosciences) for 30 minutes.
7
Lymphatic Endothelial Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
S1P was from Avanti Polar Lipids (Alabaster, AL). Recombinant murine CCL19, CCL5, CCL22, CCL2, CXCL12, CXCL10, IL-6, TNFα and IFNγ were from R&D Systems (Minneapolis, MN). Anti-VCAM-1 (clone 429), anti-ICAM-1 (clone YN1/1), anti-CD4 (GK1.5), anti-CD25 (clone PC61.5), anti-CD44 (clone IM7) and anti-TNFα were from eBioscience (San Diego, CA). Anti-human CD31, anti-human LYVE1, anti-human VEGFR3, anti-human GP38, anti-human ICAM-1 and anti-human VCAM-1 antibodies were from Biolegend (San Diego, CA). Anti-VE-cadherin was from BD PharMingen (San Diego, CA). AlexFluor-555 phalloidin was from Life Technologies (Grand Island, NY). Anti-moesin was from Cellsignaling (Danvers, MA). Anti-β-catenin, anti-collagen and anti-laminin were from Abcam (Cambridge, UK). Anti-prox1 antibody was from Bioss (Woburn, MA). Recombinant human laminin 511 was from Biolamina AB (Sundbyberg, Sweden). EIA grade gelatin was from Bio-Rad (Berkely, CA).
8
Blocking Immune Receptors in HIV T-cell Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For TLR and MHC-II blockades, PBMC was pre-treated with 10 μg of either anti-TLR-2 (eBioscience), TLR-4 (eBioscience) or MHC-II (eBioscience) blocking antibodies for 30 min. PBMC were then stimulated with FBCs from 10 HIV + ART-MSM subjects for 4 days and T cell activation was enumerated. For cytokine blockades, PBMC were treated with 10 μg of anti-TNF-α (eBioscience), IFN-γ (eBioscience) or IL-6 (eBioscience) monoclonal antibodies, then stimulated with FBCs and T cell activation was enumerated as above.
9
Isolation and Flow Cytometric Analysis of Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Splenocytes and liver mononuclear cells were isolated as described previously [16 (link)]. Flow cytometric analysis was performed using BD FACSCalibur and FACSAria III instruments. Antibodies used in this study included fluorescein isothiocyanate (FITC)-labeled anti-mouse CD49b (DX5), anti-NK1.1, anti-CD4, and anti-CD11c; Phycoerythrin (PE)-labeled anti-mouse-CD69, anti-CD107a, anti-CD44, anti-CD86, and anti-IFN-γ; PE-cyanine 5.5-labeled anti-mouse CD3e and anti-CD8; and allophycocyanin (APC)-labeled anti-mouse CD314 (NKG2D), anti-CD69, anti-CD25, anti-CD80, anti-CD62L, and anti-TNF-α, these antibodies were obtained from eBioscience (San Diego, CA, USA). FITC-B540-labeled anti-CD3, APC-cy7-labeled anti-NK1.1, PE-YG582-labeled anti-CTLA4, PE-cy7-YG780-labeled anti-TIGIT, APC-R660-labeled anti-PD-1, V450-labeled anti-LAG-3, Percpcy5.5-B695-labeled anti-Tim-3, YG780-labeled anti-Granzyme B, and B540-labeled anti-perforin were obtained from Biolegend (California,USA) and BD (New York, USA). For the analysis of intracellular molecules, cells stained with anti-CD4 and anti-CD8 antibodies were fixed using Fix/Perm Buffer (eBioscience, San Diego, CA, USA) for 30 min, incubated with anti-mouse-IFN-γ and -TNF-α for 30 min at 4 °C, and then analyzed using flow cytometry.
10
Multiparameter Flow Cytometry Panel
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!