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Tnf α percp cy5

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The TNF-α-PerCP-Cy5.5 is a fluorescently labeled antibody that binds to the Tumor Necrosis Factor alpha (TNF-α) protein. It can be used for the detection and quantification of TNF-α in flow cytometry applications.

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Lab products found in correlation

Cd4 efluor450, by Thermo Fisher Scientific (1 mentions) Granzyme b pe, by BD (1 mentions) Cd161 apc, by Miltenyi Biotec (1 mentions) Il 4 pe, by BioLegend (1 mentions) Ifn γ percp cy5, by BioLegend (1 mentions) Perforin fitc, by Thermo Fisher Scientific (1 mentions) Kaluza analysis software, by Beckman Coulter (1 mentions) Cd8 apc h7, by BD (1 mentions) Brefeldin a, by Merck Group (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Ccr7 pe cy7, by BD (1 mentions) Ifn γ pe cy7, by BD (1 mentions) α4β7 apc, by BD (1 mentions) Fix perm solution, by BD (1 mentions) Granzyme b apc, by Thermo Fisher Scientific (1 mentions) Cd56 pe, by BioLegend (1 mentions) Cd16 pe cy7, by BioLegend (1 mentions) Cd3 pe, by BioLegend (1 mentions) Il 6 apc, by BioLegend (1 mentions) Ifnγ apc cy7, by BioLegend (1 mentions)

Close spelling variants of this product

TNF-α (392 citations) PerCP-Cy5.5 (133 citations)

4 protocols using tnf α percp cy5

1

Comprehensive Cytokine Profiling of Stimulated PBMC

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Blood samples (heparin) were diluted 1:1 with RPMI and stimulated with 40 nM PMA and 2 nM Ca2+ ionophore A23187 in the presence of 3 µM brefeldin A (all Sigma) for 4 h. Subsequently, red blood cells were lysed with ammonium chloride. The remaining cells were treated with Fix/Perm reagents A and B (Invitrogen) and stained with the following antibodies: CD3-eFluor 605, CD4-eFluor450, interleukin (IL)-17-AF488, Perforin-FITC (eBioscience), CD161-APC (Miltenyi), CD8-APC-H7, Granzyme-B-PE (BD), IFN-γ-PerCP-Cy5.5, IL-4-PE, and TNF-α-PerCP-Cy5.5 (Biolegend). Samples were measured on an LSR-II flow cytometer (BD) and analyzed with Kaluza Analysis Software (Beckman Coulter).
van der Geest K.S., Kroesen B.J., Horst G., Abdulahad W.H., Brouwer E, & Boots A.M. (2018). Impact of Aging on the Frequency, Phenotype, and Function of CD161-Expressing T Cells. Frontiers in Immunology, 9, 752.
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2

Comprehensive Characterization of γδ T Cell Subsets

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γδ T cells subsets were identified using a fixable aqua blue dead cell stain (Life Technologies), and a combination of antibodies including CD3-APC-Cy7 (SP34-2), pan γδ TCR-PE (B1), CD4-Pac blue (L200) (all from BD Bioscience); Vδ2-FITC (15D;Thermofisher); and CD8-Ax700 (RPA-T8;eBioscience). CD3+ T cells were divided into Vδ1+ and Vδ2+ populations as described by Harris et al. (8 (link)) and as illustrated for a mucosal sample (Suppl Fig. 1), and were further subdivided by their CD4 and CD8 expression patterns. NKG2 receptor expression was assessed using combinations of anti-NKG2A-APC (Z99; Beckman Coulter) and anti-NKG2D-PE-Cy7 (1D11; BioLegend). For homing receptor expression and activation markers the following antibodies were used: α4β7-APC (NIH NHP Reagent Resource) and CCR7-PE-Cy7 (3D12 (BD BioScience) and CD69-Pac blue (FN50; BioLegend). For representative staining see Suppl Fig.2). For intracellular staining cells were fixed and permeabilized using a Perm/fix solution (BD Biosciences) prior to incubation with IFN-γ- PE-Cy7 (4S.B3; BDBioscience), TNF-α- PerCpCy5.5 (Mab11; BioLegend), CD107-PE-Cy5 (H4A3; BD BioScience) and Granzyme B-APC (GB12; Invitrogen). At least 50,000 CD3+ T cell events were acquired on a LSRII (BD Biosciences) and analyzed using FlowJo software version 9.8.5 (TreeStar Inc).
Tuero I., Venzon D, & Robert-Guroff M. (2016). Mucosal and systemic γδ+ T cells associated with control of SIV infection. Journal of immunology (Baltimore, Md. : 1950), 197(12), 4686-4695.
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3

Cytokine Profile of Activated PBMCs

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Peripheral blood mononuclear cells were incubated overnight with 10 ng/ml LPS (Sigma-Aldrich) in the presence of GolgiPlug (1:1,000, Becton Dickinson) after pre-stimulation with IFN-γ for 2 h. The cells were then incubated with conjugated primary antibodies in phosphate-buffered saline containing 0.5% bovine serum albumine for 30 min. The antibodies used were CD3-PE, CD14-Pacfic Blue, CD16-PE-Cy7, CD20-PE, and CD56-PE (all Biolegend) at 4°C and were incubated with EDTA for 15 min followed by incubation with FACS permeabilizing solution 2 (BD Biosciences) for 15 min. Next, conjugated antibodies to TNF-α-Percp-Cy5.5, IFN-γ-APC-Cy7, IL-1β-FITC, IL-6-APC, and IL-10-APC and their respective isotype controls (all Biolegend) were added to determine intracellular cytokine production. The cells were washed and analyzed using flow cytometry (FACSCanto II, BD Biosciences) and FACSDiva software (16 (link)).
van den Bosch T.P., Caliskan K., Kraaij M.D., Constantinescu A.A., Manintveld O.C., Leenen P.J., von der Thüsen J.H., Clahsen-van Groningen M.C., Baan C.C, & Rowshani A.T. (2017). CD16+ Monocytes and Skewed Macrophage Polarization toward M2 Type Hallmark Heart Transplant Acute Cellular Rejection. Frontiers in Immunology, 8, 346.
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4

Phenotypic Analysis of Activated T Cells

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To assess their functional phenotype, 106 T cells were activated with 50 ng/mL phorbol myristic acid and 1 µg/mL ionomycin for 30 min followed by incubation for 3 h at 37°C in the presence brefeldin A (eBioscience). Cells were fixed in fixation/permeabilization buffer (eBioscience), washed in permeabilization buffer (eBioscience), stained using anticytokine antibodies (IL-4 AF488 from eBioscience and interferon-γ AF700, IL-10 BV421, IL-17A APC/Cy7, and tumor necrosis factor (TNF)-α PerCP-Cy5.5 from BioLegend) and analyzed on an LSR II (BD Biosciences) and using FlowJo software (TreeStar Inc.).
McGinty J.W., Chow I.T., Greenbaum C., Odegard J., Kwok W.W, & James E.A. (2014). Recognition of Posttranslationally Modified GAD65 Epitopes in Subjects With Type 1 Diabetes. Diabetes, 63(9), 3033-3040.
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