Anti cd3 monoclonal antibody

Manufactured by Thermo Fisher Scientific

Sourced in Germany, United States, China

The Anti-CD3 monoclonal antibody is a laboratory tool used for the detection and analysis of T cells. It specifically binds to the CD3 complex, a group of proteins found on the surface of T cells, which plays a crucial role in T cell activation and signaling. This antibody can be used in various immunological assays and techniques to identify, quantify, and study T cell populations.

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Lab products found in correlation

Anti cd28 monoclonal antibody, by Thermo Fisher Scientific (3 mentions) Macs cd4 t cell isolation kit, by Miltenyi Biotec (2 mentions) Facsaria 3, by BD (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Neutralizing anti icosl antibody, by Thermo Fisher Scientific (1 mentions) Automated cell counter, by Countstar (1 mentions) Kbm 581 medium, by Corning (1 mentions)

Spelling variants of this product

Anti-CD3 (512 citations) Anti-CD3 antibody (84 citations) CD3 Monoclonal Antibody (5 citations) Anti-CD3 monoclonal antibody (clone OKT3; (2 citations) PE-anti mouse CD3 monoclonal antibody (mAb) (2 citations) Anti-human CD3 OKT3 monoclonal antibody (2 citations)

4 protocols using anti cd3 monoclonal antibody

Functional Characterization of T Cell Subsets in AML

Cited 1 time

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PD-1⁺CD4⁺CD25^high T cells, PD-1⁻CD4⁺CD25^high T cells, and CD4⁺CD25⁻ effector T cells were isolated from BMMNCs of 4 patients with AML by FACSAria III (Becton Dickinson, San Jose, CA, USA). The suppressive function of PD-1⁺CD4⁺ CD25^high T cells, PD-1⁻CD4⁺CD25^high T cells were determined using mixed leukocyte culture assay according to our previously reported procedure (6 (link)).
CD4⁺ T cells were selected from PBMNCs of healthy donors using MACS CD4⁺ T cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) and subsequently seeded at 5 × 10⁴ cells/well on 96-well plates coated with anti-CD3 monoclonal antibody (1 μg/ml) and stimulated with anti-CD28 monoclonal antibody (3 μg/ml) (both from eBiosciences, San Diego, CA, USA) and 20 ng/ml IL-2 for 5 days.
Full-length hPD-L1 cDNA was cloned into the lentivirus expression vector CMV-MCS-PGK-Puro, which was tranfected simultaneously with three pakaging vectors into 293FT cells to obtain virus particles. HEL cells were infected with these virus particles using pLX, and were subsequently screened with 800 μg/ml G418 for 5 days.

Dong Y., Han Y., Huang Y., Jiang S., Huang Z., Chen R., Yu Z., Yu K, & Zhang S. (2020). PD-L1 Is Expressed and Promotes the Expansion of Regulatory T Cells in Acute Myeloid Leukemia. Frontiers in Immunology, 11, 1710.

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CD4+ T Cell Activation Assay

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The CD4⁺ T cells labeled with carboxyfluorescein-succinimidyl-ester (Invitrogen, Carlsbad, CA) were cultured in the 24-well plate coated with anti-CD3 monoclonal antibody and anti-CD28 monoclonal antibody (eBioscience, San Diego, CA). After 72 h culturing, the cells were analyzed by flow cytometric analysis.

Zhao G.J., Yang X.Y., Zhang C., Dong W., Dong F.B., Zhang J., Chen X.Y., Yao R.Q., Xiao Z., Chen L.W., Yao Y.M, & Lu Z.Q. (2023). SUPPLEMENTATION WITH NICOTINAMIDE RIBOSIDE ATTENUATES T CELL EXHAUSTION AND IMPROVES SURVIVAL IN SEPSIS. Shock (Augusta, Ga.), 60(2), 238-247.

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Overexpression of ICOSL in AML cells

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Purified CD4⁺ T cells, isolated from PBMNCs of healthy donors using MACS CD4⁺ T-cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany), were seeded at 5 × 104 cells/well on 96-well plates coated with 1 μg/ml anti-CD3 monoclonal antibody (eBiosciences, San Diego, CA, USA) and stimulated with 3 μg/ml anti-CD28 monoclonal antibody (eBiosciences) and 20 ng/ml IL-2 for 5 days. Full-length hICOSL cDNA was cloned into eukaryotic expression vector GV230. HEL or HL-60 cells were transfected transiently with the positive clones of constructed recombinant plasmid GV230-hICOSL and were subsequently screened with 800 μg/ml G418 for 7 days. qPCR and western blot were performed to determine the mRNA and protein levels of ICOSL to confirm the overexpression of ICOSL. After treatment with 20 μg/ml mitomycin for 1 h, AML cells or AML overexpressed ICOSL was cocultured with CD4⁺ T cells at a ratio of 1 : 1 for 5 days in the presence or absence of 10 μg/mL neutralizing anti-ICOSL antibody (eBiosciences), with stimulation with plate-coated anti-CD3 (1 μg/ml) and soluble anti-CD28 (3 μg/ml) and IL-2 (20 ng/ml).

Zhang G., Xu Y., Zhang S, & Zhou H. (2020). The ICOSL Expression Predicts Better Prognosis for Nasopharyngeal Carcinoma via Enhancing Oncoimmunity. BioMed Research International, 2020, 9756732.

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Expansion of Primary Human NK Cells

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The fresh buffy coats (Leukocyte Source) from local blood bank (Anhui blood center) were used in this study as the source of peripheral blood mononuclear cells (PBMCs) for NK cells expansion. The PBMCs were isolated from the buffy coats by using Ficoll-Paque (Biosharp, Anhui, China). A modified feeder-free human NK cells expansion system, as described previously, was used for NK cell expansion [31 (link),32 (link)]. In brief, 6 × 10⁷ PBMCs were cultured in KBM-581 medium (Corning, 88-581-CM) supplemented with 5% of heat-inactivated autologous plasma, 1000 IU/mL rhIL-2 (Jinsili, Jiangsu, China) and anti-CD3 monoclonal antibody (eBioscience, San Diego, CA, USA), in an anti-CD16 monoclonal antibody (Beckman Coulter, Inc., Brea, CA, USA) immobilized culture flask. The cultures were continued to culture for 17–27 days at 5% CO₂, 37 °C, by adding fresh medium every 2–3 days to keep the cell density between 1.8 and 3.2 × 10⁶ cells/mL until the desired cell number was reached. Total cell numbers were counted using trypan blue by an automated cell counter (Countstar, Shanghai, China). To determine the percentage of NK cells, cells were stained for CD3 and CD56, followed by flow cytometry analysis. The final cell qualified indicators included proportion of living cells ≥90% and proportion of CD56⁺ cells ≥80%.

Chen M., Li Y., Wu Y., Xie S., Ma J., Yue J., Lv R., Tian Z., Fang F, & Xiao W. (2021). Anti-Tumor Activity of Expanded PBMC-Derived NK Cells by Feeder-Free Protocol in Ovarian Cancer. Cancers, 13(22), 5866.

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