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Cd3 pe

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CD3-PE is a fluorochrome-conjugated monoclonal antibody that recognizes the CD3 antigen. CD3 is a T cell co-receptor that is associated with the T cell receptor (TCR) and is expressed on the surface of all mature T cells. The PE (Phycoerythrin) fluorochrome allows for the detection and identification of CD3-positive cells using flow cytometry.

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Lab products found in correlation

Cd4 apc, by BioLegend (6 mentions) Cd4 apc, by Thermo Fisher Scientific (6 mentions) Ccr7 apc cy7, by BioLegend (6 mentions) Cd45ro fitc, by Miltenyi Biotec (6 mentions) Cd8a percp cy5, by Thermo Fisher Scientific (6 mentions) Cd8 apc, by BioLegend (4 mentions) Cd4 pe cy7, by BioLegend (4 mentions) Cd19 apc, by BioLegend (4 mentions) Facscanto 2, by BD (3 mentions) Cd45 fitc, by BioLegend (3 mentions) Lsrii flow cytometer, by BD (3 mentions) Cd19 bv421, by BioLegend (3 mentions) Lsrfortessa, by BD (3 mentions) B220 pe cy7, by BioLegend (3 mentions) Cd8 fitc, by BioLegend (3 mentions) Lsrfortessa ow cytometer, by BD (3 mentions) Cd8a pe cy7, by BioLegend (2 mentions) Cd45 percp, by BioLegend (2 mentions) B220 apc, by BioLegend (2 mentions) Cd11b fitc, by BioLegend (2 mentions)

Spelling variants of this product

CD3-PE-Cy7 (67 citations) PE anti-human CD3 (9 citations) Anti‐human CD3‐PE‐Cy7 (8 citations) PE/Cy7 antihuman CD3 antibody (5 citations) Anti-CD3-PE-Cy7 (clone SK-7, (4 citations) CD3 PE-Cy7 clone (3 citations) PE/Cy7 anti-human CD3 Clone HI3A (3 citations) CD3 PE/Dazzle 594 (clone 17A2 (2 citations) CD3 (clone SP34-2) PE-CF594 (2 citations) CD3 PE CF594 ( (2 citations)

53 protocols using cd3 pe

1

Murine PBMC Isolation and Staining

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Blood was collected in a tube with 3.8% w/v sodium citrate. Mouse PBMCs were purified from whole blood by Ficoll (PanEco, Moscow, Russia) density centrifugation. Murine PBMCs staining was performed using the following monoclonal antibodies: CD45-PerCP (103130; BioLegend, San Diego, CA, USA), CD3-PE (100308; BioLegend, San Diego, CA, USA), CD8a-PE/Cy7 (100722; BioLegend, San Diego, CA, USA), CD4-APC (100412; BioLegend, San Diego, CA, USA), CD19-FITC (152404; BioLegend, San Diego, CA, USA), and CD25-Pacific Blue (102022; BioLegend, San Diego, CA, USA).
Gomzikova M.O., Kletukhina S.K., Kurbangaleeva S.V., Neustroeva O.A., Vasileva O.S., Garanina E.E., Khaiboullina S.F, & Rizvanov A.A. (2020). Mesenchymal Stem Cell Derived Biocompatible Membrane Vesicles Demonstrate Immunomodulatory Activity Inhibiting Activation and proliferation of Human Mononuclear Cells. Pharmaceutics, 12(6), 577.
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2

Multiparametric Flow Cytometry Analysis

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Single cell suspension of splenocytes, lymph node, and peritoneal lavage were incubated with 10 μg anti-mouse unlabeled CD16/32 (2.4G2); stained with B220-APC or PeCy5, CD3-PE or PECy7, CD11b-FITC, or GL7-FITC (Biolegend), washed, examined on a BD Canto Flow analyzer, and data analyzed with FCS Express, v. 4.
Cooley L.F., El Shikh M.E., Li W., Keim R.C., Zhang Z., Strauss J.F., Zhang Z, & Conrad D.H. (2016). Impaired immunological synapse in sperm associated antigen 6 (SPAG6) deficient mice. Scientific Reports, 6, 25840.
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3

Immune Profiling of Murine Breast Tumors

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Brca1Co/Co;MMTV-Cre;p53+/- mice (N=5/group) with established tumors (4-5 mm in diameter) were treated for five doses (three times a week) and then tumor, spleen and mammary gland were collected and digested for flow cytometry as published previously 32 (link). Panel 1: CD45-VioGreen (Miltenyi, 3 μg/mL), Gr-1-PE (Miltenyi, 3 μg/mL), CD11b-FITC (Miltenyi, 3 μg/mL), CD19-PerCP/Cy5.5 (BioLegend, 2 μg/mL). Panel 2: CD45-VioGreen (Miltenyi, 3 μg/mL), CD4-FITC (Miltenyi, 3 μg/mL), CD3-PE (BioLegend, 2 μg/mL), CD8-APC (BioLegend, 2 μg/mL), CD25-PE/Cy7 (BioLegend, 2 μg/mL).
Zhang D., Baldwin P., Leal A.S., Carapellucci S., Sridhar S, & Liby K.T. (2019). A nano-liposome formulation of the PARP inhibitor Talazoparib enhances treatment efficacy and modulates immune cell populations in mammary tumors of BRCA-deficient mice. Theranostics, 9(21), 6224-6238.
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4

NK Cell Cytotoxicity Assay

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1 × 105 NK cells were added to tumor cell lines at an E/T ratio of 1 : 1 in 200 μl, spun down at 300 r.p.m. for 2 min and incubated in the presence of FITC-conjugated anti-CD107a antibody or isotype control antibody (BioLegend). After 30 min, 1 μg of GolgiStop (BD Biosciences) was added for an additional period of 3.5 h. As a positive control for NK cell degranulation, NK cells were incubated with 50 ng/ml PMA and 1 mM ionomycin (Sigma-Aldrich). When indicated, NK cells were incubated with 10 μg/ml of anti-NKG2D (1D11) or isotype control for 20 min before the addition to target cells. Cell suspensions were washed, stained for CD45-Pacific blue, CD3-PE and CD56-PE Cy7 (all from BioLegend) on ice in FACS buffer for 30 min, washed and briefly incubated with a 1 : 40 dilution of 7-AAD (BioLegend). Cells were measured on a FACS Canto II (BD Biosciences) and analyzed using FlowJo 9.3.2 software (Treestar).
Breunig C., Pahl J., Küblbeck M., Miller M., Antonelli D., Erdem N., Wirth C., Will R., Bott A., Cerwenka A, & Wiemann S. (2017). MicroRNA-519a-3p mediates apoptosis resistance in breast cancer cells and their escape from recognition by natural killer cells. Cell Death & Disease, 8(8), e2973-.
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5

Analyzing Immune Cells and Dendritic Cell Maturation

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To analyze immune cells by flow cytometry, spleens of mice after various treatments were collected and stained according to the manufacturer’s protocols. In brief, to analyze memory T cells, cells from tumors spleens were stained with antibodies against CD8a-APC (BioLegend, catalog no. 100712) and CD3-PE (BioLegend, catalog no. 100206). To analyze the maturation of DCs, cells from tumors spleens were stained with antibodies against CD80-APC (BioLegend, catalog no. 104714), CD86-PE (BioLegend, catalog no. 159204), CD11c-FITC (BioLegend, catalog no. 117306).
Feng Y., Liu Y., Ma X., Xu L., Ding D., Chen L., Wang Z., Qin R., Sun W, & Chen H. (2022). Intracellular marriage of bicarbonate and Mn ions as “immune ion reactors” to regulate redox homeostasis and enhanced antitumor immune responses. Journal of Nanobiotechnology, 20, 193.
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6

Analyzing Murine PBMC Subsets by Flow Cytometry

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To analyse the effect of CIMVs-IL2 on the number of murine PBMCs, newly isolated cells were stained with antibodies specific for the surface markers typical for various populations of mouse immune cells. Briefly, 5 × 105 murine PBMCs in 100 µL of PBS were stained with the following antibodies for 30 min in the absence of light at RT: CD45-PerCP (#103130; BioLegend, San Diego, CA, USA), CD3-PE (#100308; BioLegend, San Diego, CA, USA), CD8a-PE/Cy7 (#100722; BioLegend, San Diego, CA, USA), CD4-APC (#100412; BioLegend, San Diego, CA, USA). After staining, the cells were washed twice with PBS and analyzed by flow cytometry using the FACSAria III (BD Biosciences, San Jose, CA, USA) and data were analyzed using BD FACSDiva™ software version 7.0. A minimum of 20,000 events were acquired for each sample. Flow cytometry results were obtained in one replicate for each animal and summarized together (n = 5, five biological replicates).
Chulpanova D.S., Gilazieva Z.E., Kletukhina S.K., Aimaletdinov A.M., Garanina E.E., James V., Rizvanov A.A, & Solovyeva V.V. (2021). Cytochalasin B-Induced Membrane Vesicles from Human Mesenchymal Stem Cells Overexpressing IL2 Are Able to Stimulate CD8+ T-Killers to Kill Human Triple Negative Breast Cancer Cells. Biology, 10(2), 141.
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7

Comprehensive Immune Cell Profiling

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Animals were euthanized according to a standard protocol using isofluorane inhalation followed by cervical dislocation. Cardiac puncture was performed to collect blood for plasma analysis. Spleen, thymus, and mesenteric lymph nodes were collected and weighed, then single-cell suspensions were isolated from fresh tissues and cells were counted manually using a hemocytometer. Zombie Aqua (cat. #423101, dilution 1:1,000) was used to stain for dead cells. Cell-surface stains were applied and FACS analysis was performed using LSRII (Flow Cytometry Core, UAB). Primary antibodies were all from Biolegend and include: CD3 PE (cat. #100307, dilution 1:200), CD4 PacBlue (cat. #100427, dilution 1:200), CD8 PECy7 (cat. #100721, dilution 1:400), TCRγδ PerCP Cy5.5 (Cat # 118117, dilution 1:400), NK1.1 FITC (cat. #108705, dilution 1:100), Foxp3 AF647 (cat. #126407, dilution 1:100), and CD19 FITC (cat. #152403, dilution 1:200). Data were gated and analyzed using FlowJo software.
Leier A., Moore M., Liu H., Daniel M., Hyde A.M., Messiaen L., Korf B.R., Selvakumaran J., Ciszewski L., Lambert L., Foote J., Wallace M.R., Kesterson R.A., Dickson G., Popplewell L, & Wallis D. (2022). Targeted exon skipping of NF1 exon 17 as a therapeutic for neurofibromatosis type I. Molecular Therapy. Nucleic Acids, 28, 261-278.
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8

Comprehensive Flow Cytometry Analysis of Immune Cells

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For flow cytometry analysis of lymph nodes (LN), spleen (SP), tumors, and AT, the following antibodies and buffers were used: CD3-PE, CD4-BV510, CD8-PeCy7, F4/80-APC, MHCII-AF488, CD206-BV605 CCR8-BV421, CD39-PeCy7, NRP1-PE and PD1-BV711 from Biolegend; CD25-PeCy5.5 from Thermo Scientific and CD11b-VF450 from Tonbo. For intracellular analysis, we used anti-Foxp3-APC (Tonbo) and Helios-FITC (Biolegend), using the Foxp3/Transcription Factor Staining Buffer Kit (Tonbo) for fixation and permeabilization of the cells.
All cells were initially stained with viability dye Zombie-NIR (Biolegend) and samples were acquired in an Attune NxT® Acoustic Focusing Cytometer (Thermo Scientific) and flow cytometry data was analyzed using FlowJo 10.8.1 (Tree Star).
Núñez-Ruiz A., Sánchez-Brena F., López-Pacheco C., Acevedo-Domínguez N.A, & Soldevila G. (2022). Obesity modulates the immune macroenvironment associated with breast cancer development. PLoS ONE, 17(4), e0266827.
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9

Multicolor Flow Cytometry Panel

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Commercial antibodies (clones, fluorophores and sources) are as follows: CD3-FITC (145–2C11), CD62L-PE (Mel-14), CD4-PErCP/Cy5.5 (RM4–5), CD8-PerCP/Cy5.5 (53–6.7), CD8-APC (53–6.7), CD209b-APC (22D1), IgM-FITC (II/41), CD4-PE (GK1.5), CD25-AF488 (PC61.5), Streptavidin-PE, CD169-PE/Cy7 (3D6.112) (eBioscience, San Diego, CA); CD11b-FITC (M1/70), CD49d-FITC (R1–2), TCRβ-FITC (GL3), CD21/35-PE (7E9), CD23-APC (B3B4), Ly6G-PE (RB6–8C5), Ly6C-PerCP/Cy5.5 (HK1.4), CD11c-PE/Cy7 (N418), B220-PerCP/Cy5.5 (RA3–6B2), B220-APC (RA3–6B2), B220-APC/Cy7 (RA3–6B2), CD69-BV421 (H1.2F3),CD23-Biotin (B3B4), F4/80-APC (BM8), Ly6G-BV421 (1A8), CD45-Pacific Blue (30-F11), CD45-BV510 (30-F11), CD43-PE (1B11), CD24-PE/Cy7 (M1/69), IgD-Pacific Blue (11–26c.2a), CD69-PE/Cy7 (H1.2F3), IgD-PerCP/Cy5.5 (11–26c.2a), Ly51-AF647 (6C3), CD3-Pacific Blue (17A2), CD3-PE (17A2), TCRγ/δ-biotin (GL3), Streptavidin-PE/Cy7 (Biolegend, San Diego, CA); Siglec-F-PE (E50–2440) (BD Biosciences, San Jose, CA). Samples were preincubated with 1 μg Fc-block (2.4G2 hybridoma; ATCC).
Cells were acquired either on the BD Biosciences LSR Fortessa or with a BD FACScan flow cytometer with DxP multi-color upgrades by Cytek Development Inc. (Woodland Park, NJ) and analyzed using FlowJo software (FlowJo LLC, Ashland, OR).
Anaya E.P., Lin X., Todd E.M., Szasz T.P, & Morley S.C. (2021). Novel mouse model reveals that serine phosphorylation of L-plastin is essential for effective splenic clearance of pneumococcus. Journal of immunology (Baltimore, Md. : 1950), 206(9), 2135-2145.
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10

Multicolor Flow Cytometry Immunophenotyping

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Cell suspensions were labeled with anti-human monoclonal antibodies (mAb) targeting the following cell-surface markers: CD45-FITC, CD3-PE, CD4-Pe Cy7, CD8-BV421 and CD19-APC (all from Biolegend). Washing and reagent dilutions were done with FACS buffer (PBS containing 2% fetal calf serum and 0.05% sodium azide (NaN3). All acquisitions were performed on a Cyan ADP (Beckman Coulter) flow cytometer. Data were analysed with FlowJo software (Ashland, OR). Cellular debris and dead cells were excluded by their light-scattering characteristics.
Campos N., Myburgh R., Garcel A., Vautrin A., Lapasset L., Nadal E.S., Mahuteau-Betzer F., Najman R., Fornarelli P., Tantale K., Basyuk E., Séveno M., Venables J.P., Pau B., Bertrand E., Wainberg M.A., Speck R.F., Scherrer D, & Tazi J. (2015). Long lasting control of viral rebound with a new drug ABX464 targeting Rev – mediated viral RNA biogenesis. Retrovirology, 12, 30.
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