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Percp conjugated anti cd3

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PerCP-conjugated anti-CD3 is a lab equipment product that binds to the CD3 receptor on the surface of T cells. PerCP is the fluorescent label used for detection.

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Lab products found in correlation

Fitc conjugated anti cd4, by BD (3 mentions) Apc conjugated anti ifn γ, by BD (3 mentions) Apc cy7 conjugated anti cd4, by BD (3 mentions) Pe conjugated anti cd8, by BD (3 mentions) Pe conjugated anti cd69, by BD (3 mentions) Pe conjugated anti cd56, by BD (3 mentions) Apc cy7 conjugated anti cd3, by BD (3 mentions) Pe conjugated anti ly6g, by BD (2 mentions) Kaluza software, by Beckman Coulter (2 mentions) Apc h7 conjugated cd8, by BD (2 mentions) Brilliant violet 650 conjugated cd4, by BioLegend (2 mentions) Alexa 700 conjugated cd14 and cd19, by BioLegend (2 mentions) Apc conjugated anti γδ, by BioLegend (2 mentions) Dnase 1, by Thermo Fisher Scientific (2 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (2 mentions) Phytohemagglutinin (pha), by Merck Group (2 mentions) Apc conjugated anti nkp46, by BD (2 mentions) Fitc conjugated anti nkg2d, by Thermo Fisher Scientific (2 mentions) Fitc conjugated anti cd158a, by BD (2 mentions) Pe conjugated anti cd158b, by BD (2 mentions)

Spelling variants of this product

Anti-CD3 (541 citations) CD3-PerCP (82 citations) Anti-CD3-PerCP (69 citations) PerCP-Cy5.5-conjugated anti-CD3 (6 citations) PerCP-conjugated anti-mouse CD3 (3 citations) Percp)-conjugated CD3 (2 citations) PerCP-Cy5.5-conjugated anti-human CD3 (2 citations) PerCP-conjugated anti-CD3 antibody (2 citations)

20 protocols using percp conjugated anti cd3

1

Flow Cytometric Analysis of Immune Cell Subsets in ARDS

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Cells from the BALF were stained with surface markers, including PE-Cy7 conjugated anti-CD45 (BD Biosciences, CA, USA), AF488 conjugated anti-CD11b (BD Biosciences, CA, USA), and PE conjugated anti-Ly6G (BD Biosciences, CA, USA). Cells from blood and spleen were stained with various surface markers, including PE-Cy7 conjugated anti-CD45 (BD Biosciences, CA, USA), Percp-cy5.5 conjugated anti-CD3 (BD Biosciences, CA, USA), FITC conjugated anti-CD4 (BD Biosciences, CA, USA), PE conjugated anti-CD25 (BD Biosciences, CA, USA), and APC conjugated anti-CD39 (BD Biosciences, CA, USA).
Peripheral blood mononuclear cells (PBMCs) from ARDS patients and healthy donors were stained with PerCP conjugated anti-CD3 (BD Biosciences, CA, USA), FITC conjugated anti-CD4 (BD Biosciences, CA, USA), APC conjugated anti-Foxp3 (BD Biosciences, CA, USA), and PE conjugated anti-CD39 (BD Biosciences, CA, USA). This study was approved by the Jinling Hospital Ethics Review Committee and written informed consent was provided by all subjects or their legal representatives.
Chen C., Li X., Li C., Jin J., Wang D., Zhao Y., Gu Y., Chen M., Zhu S., Liu H., Lv T., Zhang F, & Song Y. (2021). CD39+ Regulatory T Cells Attenuate Lipopolysaccharide-Induced Acute Lung Injury via Autophagy and the ERK/FOS Pathway. Frontiers in Immunology, 11, 602605.
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2

Intracellular Cytokine Staining of T Cells

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For intracellular cytokine staining, T cells were treated for 2 h with brefeldin-A (Golgi plug; BD Pharmingen) at a concentration of 2 μg/mL, then harvested and washed in phosphate-buffered saline, containing 1% FBS and 0.1% NaN3 (staining buffer). They were then stained extracellularly using PerCP-conjugated anti-CD3, PE-conjugated anti-CD4, APC-Cy7-conjugated anti-CD8 monoclonal antibodies or the appropriate isotype controls for background determination, all purchased from BD Pharmingen (San Diego, CA, USA). Then, the cells were fixed and permeabilized using Cytofix/Cytoperm™ (BD Pharmingen), according to the manufacturer’s instructions, and stained with PeCy7-conjugated anti-human IFN-γ and APC-conjugated TNF-α (BD Pharmingen). Stained cells were then analyzed with flow cytometry using the Beckman Coulter Gallios equipped with Kaluza Software (Beckman Coulter, Brea, CA, USA).
Mariotti S., Venturi G., Chiantore M.V., Teloni R., De Santis R., Amendola A., Fortuna C., Marsili G., Grilli G., Lia M.S., Kiros S.T., Lagi F., Bartoloni A., Iacobino A., Cresta R., Lastilla M., Biselli R., Di Bonito P., Lista F, & Nisini R. (2024). Antibodies Induced by Smallpox Vaccination after at Least 45 Years Cross-React with and In Vitro Neutralize Mpox Virus: A Role for Polyclonal B Cell Activation?. Viruses, 16(4), 620.
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3

CFSE-based T cell proliferation assay with PBMC

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Thawed PBMC were rested for one hour, washed in 10% Human AB media (Gemini), and 3–6×106 PBMC were labeled with 1 ml of 1.25 µM 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE; Molecular Probes) for seven minutes. CFSE-labeled PBMC were incubated in 96-well, deep-well culture plates (Nunc, Roskilde, Denmark) at a density of 106 PBMC per well at a final volume of 1 ml for 7 days. In a subset of patients, CFSE-labeled PBMC were incubated with antigen in the presence of IL-10 receptor alPHA chain (IL-10Rα) blocking antibody (clone 37607; R&D Systems) or IgG1 isotype control antibody at 10 µg/mL. Antigens tested included media, phytohemagglutinin (PHA; 5 µg/mL; Sigma-Aldrich), uRBC, or PfSE at an E:T ratio of 1∶3 schizont equivalents. At day 7 cells were treated with 100 units DNase I (Invitrogen) in culture medium at 37°C for 10 min, washed, and stained with surface antibodies (PerCP–conjugated anti-CD3, APC-H7-conjugated CD8 (BD PHArmingen), Brilliant violet 650-conjugated CD4, Alexa 700-conjugated CD14 and CD19, and APC-conjugated anti-γδ (Biolegend)) before acquisition.
Jagannathan P., Eccles-James I., Bowen K., Nankya F., Auma A., Wamala S., Ebusu C., Muhindo M.K., Arinaitwe E., Briggs J., Greenhouse B., Tappero J.W., Kamya M.R., Dorsey G, & Feeney M.E. (2014). IFNγ/IL-10 Co-producing Cells Dominate the CD4 Response to Malaria in Highly Exposed Children. PLoS Pathogens, 10(1), e1003864.
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4

Cytokine-Mediated T Cell Activation

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PBMCs were initially stimulated with IL-2 (100 U/mL) for six days. Cells were then washed and restimulated with plate-bound anti-CD3/CD28 antibodies (anti-CD3, 0.2 μg/mL; anti-CD28, 0.4 μg/mL), IL-2 (100 U/mL), or HIV gag peptide (10 μg/mL), with or without anti-Tim-3/PD-1 antibodies (25 μg/mL). For the detection of IL-2 and IFN-γ production, GolgiStop was added 1 hour after restimulation, and 4 hours later cells were stained for PerCP-conjugated anti-CD3 and APC-cy7-conjugated anti-CD4 and intracellular APC-conjugated anti-IL-2 or APC-conjugated anti-IFN-γ, according to the manufacturer's recommendations (BD Biosciences). Intracellular expression of IL-2 or IFN-γ within CD4+ and CD8+ (CD3+CD4−) T cells was detected using an LSRII instrument and analyzed with FlowJo software.
Zhang Z.N., Zhu M.L., Chen Y.H., Fu Y.J., Zhang T.W., Jiang Y.J., Chu Z.X, & Shang H. (2015). Elevation of Tim-3 and PD-1 Expression on T Cells Appears Early in HIV Infection, and Differential Tim-3 and PD-1 Expression Patterns Can Be Induced by Common γ-Chain Cytokines. BioMed Research International, 2015, 916936.
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5

Multiparameter Flow Cytometry Panel

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The following antibodies and cell tracer were used staining for flow analysis: FITC-conjugated anti-CD158b (BD Biosciences, San Jose, CA), anti-IFN-γ (eBioscience, San Diego, CA); CFSE; PE-conjugated anti-IFN-γ (BD Biosciences, San Jose, CA), anti-TNF-α, anti-IL-22, anti-granzyme B (eBioscience, San Diego, CA), anti-GM-CSF (R&D Systems, Minneapolis, MN); PerCP-conjugated anti-CD3 (BD Biosciences, San Jose, CA); APC-conjugated anti-CD158a (Miltenyi Biotec, Bergisch Gladbach, Germany), anti-CD4 (BD Biosciences, San Jose, CA), anti-IL-17A (eiBoscience, San Diego, CA); strepavidin-PerCP; PE-Cy7-conjugated anti-CD56, anti-CD14 (BD Biosciences, San Jose, CA), Vioblue-conjugated anti-3DL1 (Miltenyi Biotec, Bergisch Gladbach, Germany), eFluor 650NC-conjugated anti-CD3 (ebBioscience, San Diego, CA). anti-mouse IgG κ/Negative Control Compensation Particles. The use of antibody for staining was performed per manufacturer’s instructions with proper titrations. Antibodies used for cytokine assays are IL-2, IL-6, IL-10, IL-15, IL-13, CCL-4 (MIP-1β), CCL-5, CXCL-10, CCL-2, CXCL-8, IFN-γ, TNF-α, TNF-β, granzyme B, TGF-β1 (R&D Systems, Minneapolis, MN), IL-4, IL-12, GM-CSF, and perforin (eBioscience, San Diego, CA).
Lin L., Ma C., Wei B., Aziz N., Rajalingam R., Yusung S., Erlich H.A., Trachtenberg E.A., Targan S.R., McGovern D.P., Heath J.R, & Braun J. (2014). Human Natural Killer Cells Licensed by KIR Genes Have an Altered Cytokine Program That Modifies CD4+ T Cell Function. Journal of immunology (Baltimore, Md. : 1950), 193(2), 940-949.
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6

Comprehensive Immune Cell Phenotyping by Flow Cytometry

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For surface marker staining, cells were stained in PBS supplemented with 1% FBS for 30 min on ice with the following antibodies (PerCP‐conjugated anti‐CD3, BV421‐conjugated anti‐CD4, PE‐conjugated anti‐CD8, BV570‐conjugated anti‐CD45, BV421‐conjugated anti‐F4/80, BV650‐conjugated anti‐CD86, APC‐conjugated anti‐CD206, PE‐Cy7‐conjugated anti‐CD25, FITC‐conjugated anti‐γδ TCR, APC‐Cy7‐conjugated anti‐CD11b, PE‐conjugated anti‐Ly‐6G, FITC‐conjugated anti‐CD49b, PE‐conjugated anti‐CD19, PE‐conjugated anti‐ST2 antibodies, and APC‐conjugated anti‐NKp46: BD Biosciences, San Jose, CA, USA). After fixation and permeabilization, staining with an APC‐conjugated anti‐FOXP3 antibody (eBioscience) was performed according to the manufacturer's instructions. For IL‐33 staining of primary mouse hepatocytes, cells were stained using a biotinylated anti‐IL‐33 monoclonal antibody (Enzo Life Biosciences, Raamsdonksveer, The Netherlands) and PE‐Cy7‐labeled streptavidin (BD Pharmingen, San Diego, CA, USA). All the flow cytometric data were analyzed and plotted using FlowJo software (TreeStar, Ashland, OR, USA).
Wang P., Shi B., Wang C., Wang Y., Que W., Jiang Z., Liu X., Jiang Q., Li H., Peng Z, & Zhong L. (2022). Hepatic pannexin‐1 mediates ST2+ regulatory T cells promoting resolution of inflammation in lipopolysaccharide‐induced endotoxemia. Clinical and Translational Medicine, 12(5), e849.
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7

Multicolor Flow Cytometry Analysis

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FITC-conjugated anti-CD44, PerCP-conjugated anti-CD3, PE-conjugated anti-CD8, FITC-conjugated anti-Granzyme A, anti-Granzyme B, anti-perforin and anti-IFN-γ were all purchased from BD Pharmingen. PE-conjugated anti-CD133 was purchased from eBiosciences. For cell surface staining, SMMC7721 and HepG2 cells were resuspended in 100μl staining buffer containing 10% FBS and put on ice for 20 min to block Fc receptors, then incubated with FITC-conjugated anti-CD44and PE-conjugated anti-CD133or isotype control for 30 min. The cells were then washed with 1ml ice-cold staining buffer for 2 times and centrifuged (300g) at 4°C for 5 min. The collected cells were suspended in 500μl staining buffer solution. PBMCs were incubated with SMMC7721 cells for 5 hours with brefeldin A at a final concentration of 10μg/ml, then collected and washed. As discussed above, PerCP-conjugated anti-CD3 and PE-conjugated anti-CD8 were used for surface staining. After that, cells were fixed and permeabilized, and then intracellular staining of FITC-conjugated anti-GranzymeA, Granzyme B, perforin and IFN-γ was performed. All samples after staining were evaluated using BD FACS Canto II with Diva software and analyzed using Flowjo 7.6.5.
Kang F.B., Wang L., Sun D.X., Li H.J., Li D., Wang Y, & Kang J.W. (2017). B7-H4 overexpression is essential for early hepatocellular carcinoma progression and recurrence. Oncotarget, 8(46), 80878-80888.
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8

Immunophenotyping of γδ T-cell Subsets

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Immunological phenotypes of γδ-T cells and their subpopulations were determined by flow cytometry. The following fluorophore-conjugated mouse monoclonal antibodies anti-human antibodies were used: PerCP-conjugated anti-CD3 (347344; BD Pharmingen, USA), PE-cy7-conjugated anti- TCRγδ (331222; Biolegend, USA), FITC-conjugated anti-NKG2D (11-5878; eBioscience, USA), FITC-conjugated anti-CD158a (556062; BD Pharmingen, USA), PE-conjugated anti-CD158b (559785; BD Pharmingen, USA).
Huang C., Xiang Z., Zhang Y., Li Y., Xu J., Zhang H., Zeng Y, & Tu W. (2021). NKG2D as a Cell Surface Marker on γδ-T Cells for Predicting Pregnancy Outcomes in Patients With Unexplained Repeated Implantation Failure. Frontiers in Immunology, 12, 631077.
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9

CFSE-based Lymphocyte Proliferation Assay

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Thawed PBMC were rested for one hour, washed in 10% Human AB media (Gemini), and 3-6 × 106 PBMC were labeled with 1ml of 1.25μM 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE; Molecular Probes) for seven minutes. CFSE-labeled PBMC were incubated in 96-well, deep-well culture plates (Nunc) at a density of 106 PBMC per well at a final volume of 1ml for 7 days. Antigens tested included media, phytohemagglutinin (PHA; 5 μg/mL; Sigma-Aldrich), uRBC, or PfSE at an E:T ratio of 1:3 schizont equivalents. At day 7 cells were treated with 100 units DNase I (Invitrogen) in culture medium at 37°C for 10 min, washed, and stained with surface antibodies (PerCP–conjugated anti-CD3, APC-H7-conjugated CD8 (BD PHArmingen), Brilliant violet 650-conjugated CD4, Alexa 700-conjugated CD14 and CD19, PE-conjugated anti-Vδ2, and APC-conjugated anti-γδ (Biolegend) before acquisition.
Jagannathan P., Kim C.C., Greenhouse B., Nankya F., Bowen K., Eccles-James I., Muhindo M.K., Arinaitwe E., Tappero J.W., Kamya M.R., Dorsey G, & Feeney M.E. (2014). Loss and dysfunction of Vδ2+ γδ T cells is associated with clinical tolerance to malaria. Science translational medicine, 6(251), 251ra117.
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10

Activation and Inhibition of T Cell Responses

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PBMCs were plated in round-bottom 96-microtiter plates at 500,000 cells/well in 200 μL complete RPMI 1640 containing anti-CD3 (5 μg/mL) and anti-CD28 (2 μg/mL) antibodies (BD Biosciences) or HIV gag peptide (10 μg/mL, XiAn Meilian Company), with or without anti-Tim-3 and anti-PD-1 (25 μg/mL) antibodies (Biolegend). Cells were cultured for three days, and GolgiStop was added during the last five hours of the culture. The cells were incubated with conjugated antibodies PerCP-conjugated anti-CD3, APC-cy7-conjugated anti-CD4, PE-conjugated anti-Tim-3, and FITC-conjugated anti-PD-1, and intracellular cytokine staining for APC-conjugated anti-IFN-γ (BD Biosciences Pharmingen) or APC-conjugated IL-2 (Biolegend) was carried out using the Cytofix/Cytoperm Fixation/Permeabilization Kit according to the manufacturer's instructions (BD Biosciences). Intracellular expression of IL-2 or IFN-γ within CD4+ and CD8+ (CD3+CD4−) T cells was detected using an LSRII instrument and analyzed with FlowJo software.
Zhang Z.N., Zhu M.L., Chen Y.H., Fu Y.J., Zhang T.W., Jiang Y.J., Chu Z.X, & Shang H. (2015). Elevation of Tim-3 and PD-1 Expression on T Cells Appears Early in HIV Infection, and Differential Tim-3 and PD-1 Expression Patterns Can Be Induced by Common γ-Chain Cytokines. BioMed Research International, 2015, 916936.
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