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Anti il 17a pe

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Anti-IL-17A-PE is a fluorescent-labeled monoclonal antibody that binds to interleukin-17A (IL-17A), a cytokine involved in inflammatory processes. It can be used in flow cytometry applications to detect and quantify IL-17A-expressing cells.

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Lab products found in correlation

Anti cd4 fitc, by Thermo Fisher Scientific (7 mentions) Anti foxp3 apc, by Thermo Fisher Scientific (5 mentions) Anti ifn γ pe, by Thermo Fisher Scientific (4 mentions) Anti foxp3 pe, by Thermo Fisher Scientific (4 mentions) Anti cd25 apc, by Thermo Fisher Scientific (4 mentions) Anti foxp3 fitc, by Thermo Fisher Scientific (3 mentions) Golgiplug, by BD (3 mentions) Ionomycin, by Merck Group (3 mentions) Facscalibur flow cytometer, by BD (3 mentions) Brefeldin a, by Merck Group (3 mentions) Anti ifn γ fitc, by Thermo Fisher Scientific (3 mentions) Anti ifn γ apc, by BD (2 mentions) Anti cd25 pe, by Thermo Fisher Scientific (2 mentions) Leucoperm, by Bio-Rad (2 mentions) Pe cy7 anti ifnγ, by Thermo Fisher Scientific (2 mentions) Anti rorγt pe, by Thermo Fisher Scientific (2 mentions) Anti t bet pe, by Thermo Fisher Scientific (2 mentions) Anti cd25 pe, by BD (2 mentions) Ef780, by Thermo Fisher Scientific (2 mentions) Anti ifn γ pe cy7, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

IL-17A (306 citations) Anti-IL-17A (34 citations) IL-17A-PE (29 citations) PE-conjugated anti-IL-17A (11 citations) Anti-human IL-17A-PE (8 citations) Anti-mouse IL-17A-PE (4 citations) Anti-IL-17A-PE (clone eBio17B7, (3 citations) P-phycoerythrin (PE) anti-mouse IL-17A Antibody (2 citations) PE-conjugated anti-human IL-17A (2 citations) Anti-IL-17A-PE-Cy7 (2 citations)

30 protocols using anti il 17a pe

1

Isolation and Characterization of Infiltrated Lymphocytes

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The infiltrated lymphocytes were isolated as described previously [20] (link). The allografts were cut into pieces and digested with 2 mg/ml collagenase D (Worthington Bio, Lakewood, NJ) combined with 10% fecal calf serum in RPMI 1640 media for 2 h at 37°C. The suspensions were filtered with cell strainers (40 µm, BD Biosciences, San Diego, CA). The isolated cells were stained with FITC-anti-CD4, APC-Cy7-anti-CD11b, PerCP-Cy5.5-anti-IFN-γ, eFlour660-anti-IL-13, PE-anti-IL-17A, APC-anti-Foxp3, and 7-AAD (eBioscience, San Diego, CA) according to eBioscience’s Best Protocols. The cells were assessed with a fluorescence activated cell sorter (FACS) Aria II (BD Biosciences, San Diego, CA) and analyzed using FCS Express 4 Plus (De Novo Software, Los Angeles, CA).
Huang X., Ren L., Ye P., Cheng C., Wu J., Wang S., Sun Y., Liu Z., Xie A, & Xia J. (2014). Peroxisome Proliferator-Activated Receptor γ Deficiency in T Cells Accelerates Chronic Rejection by Influencing the Differentiation of CD4+ T Cells and Alternatively Activated Macrophages. PLoS ONE, 9(11), e112953.
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2

Isolation and Characterization of Lung Immune Cells

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The left lung of individual rats was removed, cut into small pieces, digested with collagenase, and filtered through a 200-mesh sieve. After centrifugation, the collected single cell suspensions were subjected to density gradient centrifugation using Ficoll-Paque to isolate lymphocytes. Some of the collected lymphocytes were stimulated with 50 ng/mL phorbol 12-myristate 12-acetate (PMA) and 1 μg/mL ionomycin in the presence of brefeldin A for 5 h (BD Biosciences, USA). The cells were stained with FITC-anti-CD4, fixed, and permeabilized. Subsequently, the cells were stained with PE-anti-IL-17A, PE-anti-CD25, and APC-anti-Foxp3 (eBioscience, USA) and PE-anti-IL-4 or APC-anti-IFN-γ (BD Biosciences) for the analysis of the frequency of CD4+IL-17+ Th17, CD4+IFN-γ+ Th1, CD4+IL-4+ Th2, and CD4+CD25+Foxp3+ Tregs in total CD4+ T cells, respectively. The isotype rat Ig served as negative controls. After being washed, the stained cells were analyzed in flow cytometry in a FACSCalibur (BD FACSCanto II, USA) using CellQuest software.
The remaining lymphocytes were stained with PI using a DNA assay kit (KeyGEN Biotech, China). After being washed, the cells were analyzed for their cell cycling by flow cytometry.
Li X., Wang J., Cao J., Ma L, & Xu J. (2015). Immunoregulation of Bone Marrow-Derived Mesenchymal Stem Cells on the Chronic Cigarette Smoking-Induced Lung Inflammation in Rats. BioMed Research International, 2015, 932923.
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3

Multiparameter Flow Cytometry Analysis

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The following mouse antibodies were purchased from eBioscience: PE-anti-IL-17A, FITC-anti-Foxp3, APC-anti-Helios, PE-Cy7-anti-CD4 and anti-CD11c, Biotin-anti-ICOSL. The following were purchased from BD Biosciences: PE-anti-CD103, PerCP-anti-CD90.1, and PerCP-Cy5.5-anti-CD45.1. Samples were acquired on an LSRII instrument and data was analyzed using FlowJo software.
Landuyt A.E., Klocke B.J., Colvin T.B., Schoeb T.R, & Maynard C.L. (2019). ICOS-deficient regulatory T cells display normal induction of Il10 but readily downregulate expression of Foxp3. Journal of immunology (Baltimore, Md. : 1950), 202(4), 1039-1044.
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4

Comprehensive Ethmoid Sinus T Cell Analysis

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The ethmoid sinus mucosa tissues were digested with 5% Type IV collagenase into single cell suspensions, and were stimulated with phorbol 12-myristate 13-acetate (50ng/ml; Sigma-Aldrich, USA) and ionomycin (1 μg/ml; Sigma-Aldrich, USA) in the presence of Golgi Plug (1 μl/ml; BD Biosciences, USA) at 37°C for 6 h before flow cytometry. To analyze T cell subsets in sinus mucosa, cells were incubated with APC-Cy7-Fixable Viability Dye (eBioscience, USA) before staining for surface markers (FITC-anti-CD3, AF700-anti-CD8, BV650-anti-γδTCR) (All from BD Biosciences, USA). Then, cells were treated with Fixation/Permeabilization reagents (eBioscience, USA) and stained with intracellular markers (BV711-anti-IFN-γ, BV421-anti-IL-13; BD Biosciences, USA; PE-anti-IL-17A, APC-anti-FOXP3, PE-Cy7-anti-IL-22; eBioscience, USA). A BD LSRFortessa X-20 was applied for sample sorting. The data were analyzed after adjusting the fluorescence compensation with FlowJoV10.
Lu Y., Wu Y., Huang Y., Fang S., Li Y., Sun J, & Zhou H. (2021). Immunological Features of Paranasal Sinus Mucosa in Patients with Graves’ Orbitopathy. Frontiers in Endocrinology, 11, 621321.
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5

Multicolor Flow Cytometry Panel

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PerCP-Cy5.5-conjugated anti-TNFα, eFluor® 660 anti-GM-CSF, PE-Cy7 anti-IFNγ, PE anti-IL-17A, FITC-anti-CD45RA, PE-anti-CD45RO, eFluor® 450-anti-CD161 and unconjugated anti-CD28 antibodies (Abs) were from eBioscience. Brilliant Violet 605-anti-CCR6 and APC-anti-IL-10 Abs were purchased from BioLegend. RosetteSep negative-enrichment kits for human T cells, B cells and monocytes were from StemCell Technologies. Ficoll-Paque was from GE Healthcare, RPMI from Lonza and foetal bovine serum from Life Technologies. Lipopolysaccharide (E. coli, serotype 0111:B4), ionomycin and PMA were from Sigma-Aldrich and Golgiplug™ was from BD Bioscience and Leucoperm from AbD Serotec.
Bystrom J., Clanchy F.I., Taher T.E., Al-Bogami M.M., Muhammad H.A., Alzabin S., Mangat P., Jawad A.S., Williams R.O, & Mageed R.A. (2017). Response to Treatment with TNFα Inhibitors in Rheumatoid Arthritis Is Associated with High Levels of GM-CSF and GM-CSF+ T Lymphocytes. Clinical Reviews in Allergy & Immunology, 53(2), 265-276.
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6

Immunophenotyping of Immune Cell Subsets

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Cells were immunostained with the following monoclonal anti-human antibodies: FITC anti-Lin3, V450 anti-CD56, APC-H7 anti-CD45, APC anti-IFN-γ, PE anti-TNF-α, PE Annexin V, 7AAD (all from Becton Dickinson, Milan, Italy), PercpCy5.5-CD127, PE anti-TLR3, PE anti-TLR9, APC anti-TLR4, PE anti-TLR2, APC anti-ROR-γt, PE anti-ROR-γt, PECy7 anti-T-bet, PE anti-IL-17A, AlexaFluor647 anti-IL-17A (all from eBioscience), PE anti-CRTH2 (Biolegend, San Diego, CA) and PE anti-TLR7 (R&D Systems). Cells were immunostained with the following anti-mouse antibodies: FITC antiCD90.2, hematopoietic lineage cocktail efluor 450, PECy7 anti-T-bet, APC anti-ROR-γt, PE anti-TNF-αPE anti-IL-17A (all from eBioscience), APC-Cy7 anti-CD45, PE anti-ROR-γt, PECy7 anti-IFN-γ (all from Becton Dickinson). In all experiments, appropriate isotype control IgGs (Becton Dickinson and eBioscience) and fluorescence minus one controls were used. All antibodies were used at 1:100 final dilution. For intracellular immunostaining, cells were fixed and permeabilized using staining buffer set and permeabilization buffer (both from eBioscience) according to the manufacturer’s instruction. Cells were analyzed by flow cytometry (FACSverse, BD Bioscience, San Jose, CA).
Marafini I., Monteleone I., Di Fusco D., Cupi M.L., Paoluzi O.A., Colantoni A., Ortenzi A., Izzo R., Vita S., De Luca E., Sica G., Pallone F, & Monteleone G. (2015). TNF-α Producing Innate Lymphoid Cells (ILCs) Are Increased in Active Celiac Disease and Contribute to Promote Intestinal Atrophy in Mice. PLoS ONE, 10(5), e0126291.
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7

Quantification of T cell subsets in MOG-induced EAE

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To measure T cell differentiation induced by MOG35-55, splenocytes were isolated from mice on day 25 post-immunization and stimulated with 100μg/ml of MOG35-55 peptide in RPMI1640 medium containing 10% FBS for 48 h. During the last 4h of stimulation, the cells used for Th detection were exposed to 50ng/ml of phorbol-12-myristate-13-acetate (Sigma-Asdrich, USA) and 500ng/ml of ionomycin and 3μg/ml of Brefeldin A (Sigma), and cells for Treg detection were not. After that, cells were stained with FITC-anti-CD4, APC-anti-CD25, fixed, permeabilized, and stained for intracellular cytokines with PE-anti-IFN-γ, APC-anti-IL-4, PE-anti-IL-17A and PE-anti-FOXP3 for Th1, Th2, Th17 and Tregs, respectively (eBioscience, San Diego, CA, USA), followed by flow cytometry analysis on a FACS Calibur flow cytometer (BD Biosciences, San Jose, CA, USA). The cells were gated on living cells and a minimum of 104 cells were assayed. The gating strategy used is described in the following. Firstly, lymphocytes were gated by FSC and SSC. Based on above, CD4+IFN-γ+, CD4+IL-4+, CD4+IL-17+, and CD4+CD25+Foxp3+ lymphocytes were identified as Th1, Th2, Th17, and Treg cell, respectively. Data were analyzed with the Cell-Quest Software (BD Biosciences) in a blinded manner.
Jia Z., Liu J., Li B., Yi L., Wu Y., Xing J., Wang L., Wang J, & Guo L. (2022). Exosomes with FOXP3 from gene-modified dendritic cells ameliorate the development of EAE by regulating the balance of Th/Treg. International Journal of Medical Sciences, 19(8), 1265-1274.
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8

Generation and Characterization of T Cell-Specific SELENOI Knockout Mice

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Generation of T cell specific SELENOI KO using lck-Cre (distal promoter) and SELENOIfl/fl mice was previously described [21 (link)]. Experiments using mice included age/sex matched male and females 8–12 wks of age, and littermate SELENOIfl/fl mice served as WT controls. Animal protocols were approved by the University of Hawaii Institutional Animal Care and Use Committee. To block Fcγ receptors during flow cytometry, cells were preincubated with anti-CD16/32 (2.4G2; BD Pharmingen) for 15 min followed by antibody stains. BioLegend antibodies used at concentrations recommended by vendor included FITC-anti-CD3 (145–2C11), PE- or APC-anti-CD4 (GK1.5), PE-anti-IL-17 (TC11–18H10.1), PE-anti-IFNγ (XMG1.2), PE-anti-IL-10 (JES5–16E3), PE-anti-TGF-β (TW7–16B4), BV421anti-FoxP3 (MF-14); also used were PE-anti-T-bet (4B10; eBioscience); APC-anti-CD25 (PC61.5; eBioscience), and PE-anti-RORγt (AFKJS-9; Invitrogen). Dead cells were detected with BV421 viable dye or Aqua Dead Cell stain (Invitrogen/ThermoFisher). For immunohistochemistry, antibodies included rabbit polyclonal anti-CD3 (IS503, 1:200; Dako) and SMI-32 Ab (801702, 1:5000; BioLegend). For immunofluorescence, PE-anti-IL-17A was used (17B7, 1:500; Ebioscience). Myelin oligodendrocyte glycoprotein (MOG) 35–55 peptide (InSolution AMPK Inhibitor, Compound C) were purchased from Sigma.
Ma C., Hoffmann F.W., Nunes L.G., Urena F., Andrukhiv A., Gerschenson M., Pitts M.W, & Hoffmann P.R. (2022). Selenoprotein I deficiency in T cells promotes differentiation into tolerant phenotypes while decreasing Th17 pathology. Journal of leukocyte biology, 112(6), 1387-1397.
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9

Comprehensive Immune Cell Profiling

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Anti-CD3, anti-CD28, anti-CD16/32, anti-CD4-APCH7, anti-CD25-APC, PE anti-IL-17A, PECy7 anti-IFNγ, PeCy7 anti-CD45.1 were purchased from eBioscience (CA, USA). Anti-IL-4, anti-IFNγ, PE anti-α4β7, APC anti-CCR9, UV421 anti-IL-10 and APC or PE anti-CD11c were purchased from BioLegend (CA, USA). Anti-H3K4me3 and anti-H3 Abs were purchased from Abcam (Cambridge, UK). Anti-H3K27me3 and normal rabbit IgG Abs were purchased from Millipore (MA, USA)6 (link).
Doñas C., Neira J., Osorio-Barrios F., Carrasco M., Fernández D., Prado C., Loyola A., Pacheco R, & Rosemblatt M. (2021). The demethylase inhibitor GSK-J4 limits inflammatory colitis by promoting de novo synthesis of retinoic acid in dendritic cells. Scientific Reports, 11, 1342.
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10

Phenotyping of Immune Cell Subsets

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The following monoclonal antibodies were used in the present study: FITC-anti-CD4/PE-anti-CD8/Percp-anti-CD3, FITC-anti-CD4, PE-anti-CD25, APC-anti-CD127, FITC-anti-CD3/PE-anti-CD56 APC-anti-CD3, FITC-anti-CD56 (BD Pharmingen, San Diego, CA, USA), PE-anti-IL-17A and PE-anti-IFN-γ (eBioscience, ST, USA). Cells were stained according to standard procedures by incubating the antibodies at 4 °C for 30 min. Intracellular cytokine staining was performed by fixing and permeabilizing the cells with the Cytofix/Cytoperm Fixation/Permeabilization Kit (BD Pharmingen, CA, USA) according to the manufacturer’s instructions. Flow cytometry was performed on the BD canto II and the data was analyzed using BD Diva software (BD, San Diego, CA, USA).
Xu L., Cui G., Jia H., Zhu Y., Ding Y., Chen J., Lu C., Ye P., Gao H., Li L., Ma W., Lyu J, & Diao H. (2016). Decreased IL-17 during treatment of sputum smear-positive pulmonary tuberculosis due to increased regulatory T cells and IL-10. Journal of Translational Medicine, 14, 179.
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