Tnf α pe cy7

Manufactured by Thermo Fisher Scientific

The TNF-α-PE-Cy7 is a fluorochrome-conjugated antibody that binds to the Tumor Necrosis Factor alpha (TNF-α) protein. It is commonly used in flow cytometry applications to detect and quantify the expression of TNF-α in various cell types.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

TNF-α (2204 citations) TNF-α-PE (12 citations) Anti-TNF-α-PE-Cy7 (7 citations) TNF-PE-Cy7 (2 citations)

9 protocols using tnf α pe cy7

Ex vivo Analysis of Cytokine Responses

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Unstimulated BAL cells were stained ex vivo with anti-CD3-PE-Cy7 and CCR5-PE (both from BD) and with CD4-PE-Cy5.5 and CD8-Qdot705 (both from Invitrogen). Freshly stimulated BAL cells and stimulated cryopreserved blood cells were washed and stained with anti-CD4-PE-Cy5.5 and CD8-Qdot705 (both from BD), permeablized, and stained intracellularly with CD3-APC-H7, IFN-γ-Alexa700, and interleukin 2 (IL-2)-APC (all from BD) and with tumor necrosis factor α (TNF-α)-PE-Cy7 (eBiosciences). Cells were acquired on a BD Fortessa, using FACSDiva software, and data were analyzed using FlowJo (TreeStar) and Pestle and Spice [25 (link)]. A positive cytokine response was defined as a level that was twice the background level, a net response of >0.05%, and an event cutoff of 10 events, and all data are reported after subtraction of the background level.

Bunjun R., Riou C., Soares A.P., Thawer N., Müller T.L., Kiravu A., Ginbot Z., Oni T., Goliath R., Kalsdorf B., von Groote-Bidlingmaier F., Hanekom W., Walzl G., Wilkinson R.J, & Burgers W.A. (2017). Effect of HIV on the Frequency and Number of Mycobacterium tuberculosis–Specific CD4+ T Cells in Blood and Airways During Latent M. tuberculosis Infection. The Journal of Infectious Diseases, 216(12), 1550-1560.

+ Open protocol

+ Expand

Ex vivo intracellular cytokine staining

Check if the same lab product or an alternative is used in the 5 most similar protocols

For intracellular cytokine staining, an ex vivo brefeldin A protocol was followed. Prior to staining, brain leukocytes were incubated with BFA (10 μg/mL, Sigma) in 1 mL complete RPMI for 2 h at 37 °C (5% CO₂). Afterward, cells were resuspended in Fc Block, stained for surface antigens and washed in 100 μL of fixation/permeabilization solution (BD Biosciences) for 20 min. Cells were then washed twice in 300 μL permeabilization/wash buffer (BD Biosciences), resuspended in an intracellular antibody cocktail (0.25 μg for each antibody, 1:100 dilution) containing TNFα‐PE‐Cy7 (eBioscience) and IL‐1β‐PE (eBioscience), IL‐10‐APC and IL‐4‐PerCP‐Cy5.5 (BioLegend) and subsequently fixed.

Al Mamun A., Chauhan A., Yu H., Xu Y., Sharmeen R, & Liu F. (2018). Interferon regulatory factor 4/5 signaling impacts on microglial activation after ischemic stroke in mice. The European Journal of Neuroscience, 47(2), 140-149.

+ Open protocol

+ Expand

Multicolor Flow Cytometry Immunophenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following monoclonal antibodies were used and purchased from eBioscience: mouse anti-human α-CD3-Cyanin-7-allophycocyanin (APC-Cy7; clone OKT3); α-CD4-Pacific Blue (PB; clone RPA-T4); α-CD8-fluorescein isothiocyanate (FITC; clone SK1); α-CD25-Cyanin-7-phycoerytherin (PE-Cy7; clone BC96); and rat anti-mouse α-CD19-PE (clone 1D3); α-CD3-FITC (clone 17AD); α-CD90.1- Peridinin-chlorophyll protein-Cy5.5 (PerCP-Cy5.5) or –FITC (clone HIS51); α-CD8-PB or –PE or –V500 (clone 53-6.7); α-CD27-PE-Cy7 (clone LG.7F9); α-CD44-FITC or –PerCP-Cy5.5 (clone IM7); α-CD62L-APC-Cy7 (clone MEL-14); α-CD127-FITC (clone A7R34); α-KLRG1 (clone 2F1); INF-γ-FITC (clone XMG1.2); MIP-1α-PE (clone DNT3CC); TNF-α-PE-Cy7 (cloneMP6-XT22); and IL-2-APC (clone JES6-5H4) . The following monoclonal antibodies used to identify naïve and memory T cell subsets were purchased from BD Pharmingen: mouse anti-human α-CD27-PE (clone M-T271) and α-CD45RO-FITC (clone UCHL1). Apoptosis was assessed by FACS analysis of annexin V and 7AAD-stained cells (BD Pharmingen) as previously described (23 ). MitoTracker Red, MitoTracker Green, and LysoTracker Green were used and purchased from LifeTechnologies (Gaithersburg, MD).

McIver Z.A., Grayson J.M., Coe B., Hill J.E., Schamerhorn G.A., Ohulchanskyy T.Y., Linder M.K., Davies K.S., Weiner R.S, & Detty M.R. (2016). Targeting T cell Bioenergetics by Modulating P-glycoprotein Selectively Depletes Alloreactive T cells to Prevent GVHD. Journal of immunology (Baltimore, Md. : 1950), 197(5), 1631-1641.

+ Open protocol

+ Expand

Cytokine Production and Degranulation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

For analysis of cytokine production and CD107a degranulation, PBMCs were resuspended in RPMI containing 100 U/ml penicillin and 100 μg/ml streptomycin, 2 mM L-glutamine, and 10% FCS and incubated with monensin (1.25 μg/ml and anti-CD107a FITC (BD Biosciences, clone H4A3) for 4 hours. Following incubation, cells were stained with the phenotyping antibodies listed above. They were then fixed (2% PFA), permeabilized (1% Saponin), and incubated with the monoclonal antibodies TNF-α-PE-Cy7 (Ebioscience, clone MAb11) and IFN-γ-AF700 (Biolegend, clone 45.B3) for 20 minutes at room temperature. Cells were analyzed using an LSR-II flow cytometer (BD Biosciences).

Inman C.F., Eldershaw S.A., Croudace J.E., Davies N.J., Sharma-Oates A., Rai T., Pearce H., Sirovica M., Chan Y.L., Verma K., Zuo J., Nagra S., Kinsella F., Nunnick J., Amel-Kashipaz R., Craddock C., Malladi R, & Moss P. (2018). Unique features and clinical importance of acute alloreactive immune responses. JCI Insight, 3(10), e97219.

+ Open protocol

+ Expand

Intracellular Cytokine Profiling of Brain Leukocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

For intracellular cytokine staining, an ex vivo brefeldin A protocol was followed. Prior to staining, brain leukocytes were incubated with BFA (10 μg/mL, Sigma) in 1mL complete RPMI for 2h at 37 °C (5 % CO₂). Afterward, cells were re-suspended in Fc Block, stained for surface antigens and washed in 100μL of fixation/permeabilization solution (BD Biosciences) for 20 minutes. Cells were then washed twice in 300μL permeabilization/wash buffer (BD Biosciences), re-suspended in an intracellular antibody cocktail (0.25μg for each antibody, 1:100 dilution) containing TNFα-PE-Cy7 (eBioscience) and IL-1β-PE (eBioscience), IL-10-APC and IL-4-PerCP-Cy5.5 (BioLegend) and subsequently fixed.

+ Open protocol

+ Expand

Cytokine and Degranulation Assay for T Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Following identification of CMV positive and negative donors, 2 × 10⁶ cells were incubated at 37°C for 5 h with either PMA and ionomycin, CMV peptide mix (10 (link)) or EBV peptide mix at 10 μg/ml (Supplementary Table 6), along with protein cocktail inhibitor mix (eBiosciences). Live/dead red dye (Invitrogen), anti-CD3 APC-Cy7 (Biolegend) anti-CD8 Amcyan (BD Biosciences), were then applied before fixation and permeabilisation and IFN-γ AF700 (Biolegend) and TNF-α PE-Cy7 staining (eBioscience). Example flow plots can be found in supplementary (Figure S1). For assessing immune cell activation and cytotoxic degranulation, 2 × 10⁶ cells were stimulated with either the above peptide mixes overnight at 37°C or a cell stimulation cocktail (Invitrogen) for 5 h. At the time of stimulation, CD107a FITC (Biolegend), along with brefeldin A and monensin was incorporated into the stimulation panel (example staining of CD107a can be found in supplementary).

Parry H.M., Mirajkar N., Cutmore N., Zuo J., Long H., Kwok M., Oldrieve C., Hudson C., Stankovic T., Paneesha S., Kelly M., Begum J., McSkeane T., Pratt G, & Moss P. (2019). Long-Term Ibrutinib Therapy Reverses CD8+ T Cell Exhaustion in B Cell Chronic Lymphocytic Leukaemia. Frontiers in Immunology, 10, 2832.

+ Open protocol

+ Expand

Ex Vivo Intracellular Cytokine Staining

Check if the same lab product or an alternative is used in the 5 most similar protocols

For intracellular cytokine staining, an ex vivo brefeldin A protocol was followed. Prior to staining, brain leukocytes (1×10⁶ cells) were incubated with BFA (10 μg/mL, Sigma) in 1mL complete RPMI for 2h at 37 °C (5 % CO2). Afterward, cells were re-suspended in Fc Block, stained for surface antigens and washed in 100μL of fixation/permeabilization solution (BD Biosciences) for 20 minutes. Cells were then washed twice in 300μL permeabilization/wash buffer (BD Biosciences), re-suspended in an intracellular antibody cocktail (0.25μg for each antibody, 1:100 dilution) containing TNFα-PE-Cy7 (eBioscience) and IL-1β-PE (eBioscience), IL-10-APC and IL-4-PerCP-Cy5.5 (BioLegend), and subsequently fixed. Data were acquired on a CytoFLEX (Beckman Coulter) and analyzed with FlowJo (Treestar Inc.).

Mamun A.A., Yu H., Mirza M.A., Romana S., McCullough L.D, & Liu F. (2018). Myeloid Cell IRF4 signaling protects neonatal brains from hypoxic ischemic encephalopathy. Neurochemistry international, 127, 148-157.

+ Open protocol

+ Expand

Multicolor Flow Cytometry for Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

The antibodies used were: CD3-PB (clone 17A2), CD3-AF700 (clone 17A2), CD4-Percp Cy5.5 (clone RM4-4), CD44-APC Cy7 (clone IM7), and CD45-PB (clone 30-F11) from BioLegend; CD4-Alexa Fluor 647 (clone RM4-5), CD8-FITC (clone 53-6.7), IFN-γ-APC-Cy7 (clone XMG1.2), and IL-2-PE (clone JES6-5H4) from BD Biosciences; and CD4-FITC (clone GK1.5), CD8a-Percp Cy5.5 (clone 53-6.7), CD11b-Percp Cy5.5 (clone M1/70), CD14-Percp Cy5.5 (clone Sa2-8), CD16/32-Percp Cy5.5 (clone 93), CD19-Percp Cy5.5 (clone 1D3), CD103-PE (clone 2E7), KLRG1-PE Cy7 (clone 2F1), CXCR3-PE (clone CXCR3-173), and TNF-α-PE Cy7 (clone MP6-XT22) from eBioscience.

Hu Z., Wong K.W., Zhao H.M., Wen H.L., Ji P., Ma H., Wu K., Lu S.H., Li F., Li Z.M., Shu T., Xu J.Q., Lowrie D.B, & Fan X.Y. (2017). Sendai Virus Mucosal Vaccination Establishes Lung-Resident Memory CD8 T Cell Immunity and Boosts BCG-Primed Protection against TB in Mice. Molecular Therapy, 25(5), 1222-1233.

+ Open protocol

+ Expand

Multiparameter Flow Cytometry Panel

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following reagents were used: PD-1^FITC, CTLA-4^APC, CD4^{FITC/APC-H7/V500}, CD14^PerCP, CD19^PerCP, CD3^PerCP, Via Probe, Monensin, functional grade anti-CTLA-4 (BD Biosciences); CD244^FITC, PD-1^APC, CD127^APC-Cy7, IL-2^FITC, IFN-γ^PE, TNF-α^PE-Cy7, anti-PD-L1/2 and anti-IgG1 isotype control (eBioscience); TIM-3^APC (R&D Systems); CCR7^FITC, CD57^PacificBlue, CD45RA^PacificBlue, functional grade anti-TIM-3 (Biolegend). Micro-Beads^anti-PE (Miltenyi). KLRG1^{AlexaFluor488} (clone: 13F12F2) was generated as described [14] (link).

Raziorrouh B., Heeg M., Kurktschiev P., Schraut W., Zachoval R., Wendtner C., Wächtler M., Spannagl M., Denk G., Ulsenheimer A., Bengsch B., Pircher H., Diepolder H.M., Grüner N.H, & Jung M.C. (2014). Inhibitory Phenotype of HBV-Specific CD4+ T-Cells Is Characterized by High PD-1 Expression but Absent Coregulation of Multiple Inhibitory Molecules. PLoS ONE, 9(8), e105703.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!