Anti cd3 apc

Manufactured by BioLegend

Sourced in United States, United Kingdom

Anti-CD3-APC is a monoclonal antibody that binds to the CD3 antigen expressed on the surface of T cells. It is conjugated with the fluorescent dye Allophycocyanin (APC), which can be detected using flow cytometry.

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Lab products found in correlation

Anti cd4 fitc, by BioLegend (15 mentions) Anti cd8 pe, by BioLegend (8 mentions) Anti cd8 fitc, by BioLegend (7 mentions) Anti cd4 pe, by BioLegend (5 mentions) Anti cd11c apc, by BioLegend (4 mentions) Anti cd45 fitc, by BioLegend (4 mentions) Mitomycin c, by Merck Group (4 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (4 mentions) Anti cd3 pe, by BioLegend (3 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (3 mentions) Facsaria 2, by BD (3 mentions) Facsdiva software, by BD (3 mentions) Anti cd45 pacific blue, by BioLegend (3 mentions) Anti ter119 fitc, by BioLegend (3 mentions) Anti sca1 pacific blue, by BioLegend (3 mentions) Anti pdgfrα pe, by BioLegend (3 mentions) Ack solution, by Thermo Fisher Scientific (3 mentions) Anti cd31 fitc, by BioLegend (3 mentions) Facscanto 2, by BD (3 mentions) Anti cd80 pe, by BioLegend (3 mentions)

Close spelling variants of this product

APC-Cy7-conjugated anti-mouse CD3 Anti-CD3-APC (clone 145-2C11, APC)-conjugated anti-human CD3 antibody APC-conjugated anti-human CD3 APC/Cy7 anti-human CD3 antibody APC-conjugated anti-CD3 antibody APC-Cy7-conjugated anti-human CD3 Anti-CD3 APC/Cyanine7 APC-conjugated anti-mouse CD3 antibody APC/Cyanine7 anti-mouse CD3 antibody

66 protocols using anti cd3 apc

Comprehensive Immune Cell Profiling of CNS

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Cells were suspended in 1 ml of FACS buffer (PBS with 2% FBS). After Fc receptor blocking with antimouse CD16/32 (BioLegend), cells were incubated with a mix of antibodies specific to analysis at 1:200 dilution each for 20 min, and dead cell marker 7-aminoactinomycin D (5 µl/sample) or DAPI (1 µl/sample) was added right before acquisition on a BD FACSCanto II device. Data were analyzed using BD FACSDiva software. Cell counts were reported to initial cell number determined after Percoll density gradient purification.
The antibody panel for CNS-infiltrating cells was as follows: Alexa Fluor 488 anti-CD45, APC anti-CD3, PE anti-CD11b, PE–cyanine 7 anti-F4/80, and 7- aminoactinomycin D (BioLegend). The antibody panel for RNAseq was as follows: Alexa Fluor 488 anti-CD45, APC anti-CD3, PE anti-CD11b, and DAPI (BioLegend).

Mimouna S., Rollins D.A., Shibu G., Tharmalingam B., Deochand D.K., Chen X., Oliver D., Chinenov Y, & Rogatsky I. (2020). Transcription cofactor GRIP1 differentially affects myeloid cell–driven neuroinflammation and response to IFN-β therapy. The Journal of Experimental Medicine, 218(1), e20192386.

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Profiling Immune Cell Composition in Mouse Lungs

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GFP-expressing AAV or lentiviral vectors were administrated i.n. into hACE2 K18 Tg (AAV) or SAMHD1 KO mice (lentivirus). At 3-dpi, the lungs were homogenized in ACK buffer and the cells were disaggregated by a 30-min treatment with 1.5 mg/mL collagenase and 0.1 mg/mL DNase followed by passage through a 100-μm mesh. The cells were blocked with anti-CD16/CD32 (RRID:AB_1574975) and stained with Alexa 700-anti-CD45 (RRID:AB_493715), PerCP-Cy5.5-anti-F4/80 (RRID:AB_893496), APC-Cy7-SiglecF (RRID:AB_2904295), PE-Cy7-anti-CD11c (RRID:AB_493569), PE-Cy7-anti-CD19 (RRID:AB_313655), APC-anti-CD3 (RRID:AB_2561456), Pacblue-anti-CD11b (RRID:AB_755985), PE-Cy5.5-anti-CD62L (RRID:AB_313097), APC-anti-CD14 (RRID:AB_940574) and PE-Ly6C/Ly6G (Gr1) (RRID:AB_313372) (BioLegend) and analyzed on a Beckman CytoFLEX flow cytometer using with FlowJo software. Cell types were classified as epithelial (CD45−), alveolar macrophages (CD45+, F4/80+, SiglecF+), interstitial macrophages (CD45+, F4/80+, SiglecF−), DCs (CD45+, F4/80−, CD11c+), T cells (CD45+, CD3+), B cells (CD45+, CD19+), monocytes (CD45+, CD11b+, CD14+) and neutrophils (CD45+, CD62L+, Ly6C/Ly6G+). LY-CoV1404 was obtained from discarded vials of Bebtelovimab.

Tada T., Minnee J, & Landau N.R. (2023). Vectored immunoprophylaxis and treatment of SARS-CoV-2 infection in a preclinical model. Proceedings of the National Academy of Sciences of the United States of America, 120(23), e2303509120.

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Immune Response Modulation Assay

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A647 anti-MICA/B, APC anti-CD3, FITC anti-CD56, APC anti-HLA-E, PE anti-HLA-A/B/C, blocking anti-NKG2D and all isotype control were from Biolegend. PE anti-MICA, -MICB, ULBP1, -ULBP2 and -ULBP3 were from R&D systems. FITC anti-pan γδTCR was from Beckman Coulter. PE anti-CD107a was from BD. Cetuximab (Erbitux) was from Merck. Recombinant human EGF was from Calbiochem. All cell culture media and reagents, Alexa 488 goat anti-rabbit IgG, and Prolongold were from Invitrogen. Chemical inhibitors and rabbit polyclonal anti-AUF1 and anti-phosphoAUF1 were from Sigma-Aldrich. The Dual Luciferase reporter system was from Promega.

Vantourout P., Willcox C., Turner A., Swanson C., Haque Y., Sobolev O., Grigoriadis A., Tutt A, & Hayday A. (2014). Immunological Visibility: Posttranscriptional Regulation of Human NKG2D Ligands by the EGF Receptor Pathway. Science translational medicine, 6(231), 231ra49.

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In Vivo Leukocyte Trafficking Dynamics

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Two hundred microliters of DSM or ES-DSM (concentrations of DOX and SCH were 300 and 50 μg/mL, respectively) was injected into the mice via the tail vein, and at 2, 8, and 24 h after injection, the leukocytes of treated mice were isolated by the mouse peripheral blood leukocyte separation kit according to the manufacturer’s instructions (Solarbio, China). The DOX fluorescence on the obtained leukocytes was analyzed by flow cytometry (ACEA NovoCyte, USA) and confocal laser scanning microscope (CLSM) (Leica SP8, Germany). In addition, leukocytes were isolated 24 h after ES-DSM injection, the T lymphocytes and neutrophils were labeled by APC-anti-CD3 (Cat#100235, 1:40) or CD16 (Cat#158005, 1:40) antibody (BioLegend, USA), respectively, then observed by CLSM.

Qi J., Jin F., You Y., Du Y., Liu D., Xu X., Wang J., Zhu L., Chen M., Shu G., Wu L., Ji J, & Du Y. (2021). Synergistic effect of tumor chemo-immunotherapy induced by leukocyte-hitchhiking thermal-sensitive micelles. Nature Communications, 12, 4755.

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Immunological Characterization of BMDM

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Apoptosis of BMDM were determined by using FITC Annexin V apoptosis detection kit with Propidium Iodide (PI) (BD biosciences), MHC-II expression of BMDM in response to rRv1768 stimulation was measured by PerCP/Cy5.5 anti-MHC-II (Biolegend, CA, USA). Cytokines production in culture supernatant of rRv1768 stimulated BMDM was determined using mouse Th1/Th2/Th17 cytometric-beads array kit (BD PharMingen, USA), according to the manufacturer's protocol. Intracellular IFN-γ expression of CD3⁺CD4⁺ T cells was determined by staining with APC-anti-CD3, FITC-anti-CD4, PE-anti-IFN-γ (Biolegend, CA, USA). All procedures were performed as we previously described (Tang et al., 2017 (link); Yuan et al., 2019 (link)) and analyzed on BD Accuri C6 Flow cytometer (BD Biosciences).

Yuan C.H., Zhang S., Xiang F., Gong H., Wang Q., Chen Y, & Luo W. (2019). Secreted Rv1768 From RD14 of Mycobacterium tuberculosis Activates Macrophages and Induces a Strong IFN-γ-Releasing of CD4+ T Cells. Frontiers in Cellular and Infection Microbiology, 9, 341.

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Multicolor Flow Cytometry Gating Strategy

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Multicolor flow cytometry gating strategy was applied to analyze various cell populations simultaneously. Expanded cells were co-stained with 6 different antibodies and analyzed with LSRII flow cytometer (BD Bioscience, San Jose, CA). Briefly, cells were re-suspended in 100 μl MACS buffer (Miltenyi Biotec, Auburn, CA) and stained with antibody cocktail in the dark at 4°C for 15 minutes. Washing twice with MACS buffer was performed before and after cell staining. The antibody cocktail contains: FITC-conjugated anti-Vδ2 TCR (Biolegend, San Jose, CA), APC-conjugated anti-CD3 (Biolegend), PE conjugated anti-CD4 (BD), PE-CyC7-conjugated anti-CD56, APC-CyC7-conjugated anti-CD16, V450-conjugated anti-CD8a Beckman Coulter (Fullerton, CA). The antibody cocktail used for Treg analysis contains CD4-PE, CD25-APC (BD Bioscience, San Jose, CA), and CD127-FITC (eBioscience) to detect CD4+CD25+CD127- Treg population. The gating strategies used in the current study are included in S1 and S2 Figs. Cell populations were also analyzed with one- or two-color flow cytometry in Accuri C6 flow cytometer (BD). FITC-anti-Vδ2 TCR, APC-anti-CD3, and PE-anti-CD3, and APC-anti-CD56 (Biolegend) were used.

Du S.H., Li Z., Chen C., Tan W.K., Chi Z., Kwang T.W., Xu X.H, & Wang S. (2016). Co-Expansion of Cytokine-Induced Killer Cells and Vγ9Vδ2 T Cells for CAR T-Cell Therapy. PLoS ONE, 11(9), e0161820.

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T Cell Activation Regulation by Arginase

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Human T cells were stained with CellTrace CSFE staining solution (Thermo Fisher Scientific, Waltham, MA, USA). Cells were then incubated with Invitrogen Dynabeads Human CD3/CD28 T Cell Activator (Thermo Fisher Scientific, Waltham, MA, USA) and IL-2 (Thermo Fisher Scientific, Waltham, MA, USA) for 96 h in the presence or absence of arginase-rich supernatant. Different percentages of supernatants from arginase-transfected 293T cells were used. Flow cytometer was used for gating for CD4 and CD8 T cells, as well as 488 nm excitation and emission for the CFSE fluorescence. Antibodies used for T cell gating included allophycocyanin (APC) anti-CD3 (BioLegend, San Diego, CA, USA), phycoerythrin (PE) CD4 (BioLegend, San Diego, CA, USA), and APC-Cy7 anti-CD8 (BioLegend, San Diego, CA, USA). The experiment was performed in triplicates.

Khoshnejad M., Perales-Puchalt A., Dia Y., Xiao P., Patel A., Xu Z., Zhu X., Yun K., Baboo I., Qureshi R., Humeau L., Muthumani K, & Weiner D.B. (2020). Synthetic DNA Delivery of an Engineered Arginase Enzyme Can Modulate Specific Immunity In Vivo. Molecular Therapy. Methods & Clinical Development, 18, 652-663.

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Identification of iNKT Cell Subsets

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The following reagents were used to stain 2x10⁶ PBMCs for ex vivo iNKT analysis: PE-conjugated CD1d-αGC tetramers, FITC anti-Vα24 (Clone C15, Beckman Coulter, High Wycombe, UK) and APC anti-CD3 (Clone SP34-2, BD Biosciences, Oxford, UK). In vitro αGC expanded iNKT cells were stained with the following reagents: PE-Cy7 anti-CD8 (Clone SK1, Biolegend, London, UK), PerCP-Cy5.5 anti-CD4 (Clone OKT4, Biolegend, London, UK), PE-conjugated CD1d-αGC tetramers, FITC anti-Vα24 and APC anti-CD3 to identify iNKT subsets. Propidium iodide (Sigma-Aldrich, Gillingham, UK) was added to identify live cells, before acquisition on either a FACSCalibur or a FACSAria II (BD Biosciences, Oxford, UK). Analysis was performed using Cell Quest (version 0.3.bfab, BD Biosciences, San Jose, US) or FloJo (version 9.7.6, Treestar, Ashland, OR, US) software; and cells were gated on live, CD3⁺ lymphocytes.

Chancellor A., White A., Tocheva A.S., Fenn J.R., Dennis M., Tezera L., Singhania A., Elliott T., Tebruegge M., Elkington P., Gadola S., Sharpe S, & Mansour S. (2017). Quantitative and qualitative iNKT repertoire associations with disease susceptibility and outcome in macaque tuberculosis infection. Tuberculosis (Edinburgh, Scotland), 105, 86-95.

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Multimodal Nanomaterial-Based Vaccine Delivery

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Tetraethyl orthosilicate (TEOS), ethanol, ammonia, potassium permanganate (KMnO₄), and sodium carbonate (Na₂CO₃) were purchased from Macklin (China). Ovalbumin (OVA) was obtained from Sigma (USA). OVA-Cy5.5 was obtained from QIYUE BIOLOGY(China). DNA single strands A₅₀ and T₂₀ were brought from Tsingke Biotech (China). Enhanced CCK-8 kit, Lyso-Tracker green fluorescent dye, 4’,6-diamidino-2-phenylindole (DAPI) and BCA protein assay kit were purchased from Beyotime (China). Roswell Park memorial institute (RPMI) 1640 culture medium, dulbecco’s modified eagle medium (DMEM), and fetal bovine serum (FBS) brought from Procell (China). Anti-CD11c-APC, anti-MHC II-PE, anti-MHC I-PE, anti-CD80-Cy5.5, anti-CD86-FITC, FITC-anti-CD4, Cy5.5-anti-CD8a, PE-anti-CD44, APC-anti-CD62L and APC-anti-CD3 antibodies were supplied by BioLegend (USA). Enzyme-linked immunosorbent assay (ELISA) Kits for INF-γ, TNF-α, IL-4, and IL-6 were also purchased from BioLegend (USA). In addition, all animal experiments were determined eligible for the study and were approved by the Ethics Committee of Jinan University.

Zha Y., Fu L., Liu Z., Lin J, & Huang L. (2024). Construction of lymph nodes-targeting tumor vaccines by using the principle of DNA base complementary pairing to enhance anti-tumor cellular immune response. Journal of Nanobiotechnology, 22, 230.

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Immunomodulatory Effects of SM Compound

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SM was synthesized by Beijing Institute of Pharmacology and Toxicology and formulated into the desired concentration with 1,2-propanediol solution (Sigma, St. Louis, MO, USA) before use. ³H-TdR was obtained from the China Institute of Atomic Energy (Beijing, China). RPMI 1640 and calf serum were from Gibco (Carlsbad, CA, USA). APC-anti-CD3, PE-anti-CD4, and FITC-anti-CD8 were from BioLegend (San Diego CA, USA). The Luminex mouse cytokine kit was from Millipore (San Diego, CA, USA). The Annexin V-FITC apoptosis kit was from Becton, Dickinson and Company (Franklin Lakes, NJ, USA). All other chemicals were obtained from Sigma (St. Louis, MO, USA) unless otherwise indicated.

Mei Y.Z., Zhang X.R., Jiang N., Cheng J.P., Liu F., Zheng P., Zhou W.X, & Zhang Y.X. (2014). The injury progression of T lymphocytes in a mouse model with subcutaneous injection of a high dose of sulfur mustard. Military Medical Research, 1, 28.

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