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Cd3 alexa fluor 700

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The CD3 Alexa Fluor 700 is a fluorescently labeled antibody that binds to the CD3 protein complex on the surface of T cells. It is designed for use in flow cytometry applications to identify and quantify T cell populations.

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Lab products found in correlation

Cd4 v500, by BD (3 mentions) B220 percp cy5, by Thermo Fisher Scientific (2 mentions) Cd11c fitc, by Thermo Fisher Scientific (2 mentions) Cd44 efluor450, by Thermo Fisher Scientific (2 mentions) Cd8 efluor605, by Thermo Fisher Scientific (2 mentions) Gr 1 pecy7, by Thermo Fisher Scientific (2 mentions) Biotinylated antibodies against cd3, by Thermo Fisher Scientific (2 mentions) B6 rag mice, by Jackson ImmunoResearch (2 mentions) Anti cd16 32 2.4g2 and normal mouse sera, by Jackson ImmunoResearch (2 mentions) Cd11b pe tr, by Thermo Fisher Scientific (2 mentions) Cd4 fitc, by Thermo Fisher Scientific (2 mentions) Protein transport inhibitor mixture, by Thermo Fisher Scientific (1 mentions) Ifnγ apc cy7, by Thermo Fisher Scientific (1 mentions) Dissociation enzyme mix, by Miltenyi Biotec (1 mentions) Cd8a buv 737, by BD (1 mentions) Cd25 apccy7, by Thermo Fisher Scientific (1 mentions) Rorγt percp cy5, by BD (1 mentions) Fas pe cf594, by BD (1 mentions) Cd45ro pe cf594, by BD (1 mentions) Fasl pe, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Alexa Fluor 700 (33 citations) Alexa Fluors (21 citations) Anti-CD3 Alexa Fluor 700 (6 citations) Anti-CD3-Alexa Fluor 700 (UC-HT1), (3 citations) Anti-human CD3-Alexa Fluor 700 (2 citations) Anti-human CD3-Alexa Fluor 700 (OKT3; (2 citations)

7 protocols using cd3 alexa fluor 700

1

Adoptive Transfer of Mpzl3-Deficient Lymphocytes

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Bone marrow was flushed out of femurs and tibias of Mpzl3 −/− or +/+ littermates (6 weeks old). The bone marrow was labeled using biotinylated antibodies against CD3, CD4, CD8 and B220 (eBioscience, San Diego, CA). Streptavidin-labeled microbeads were then used to magnetically deplete the biotinylated lymphocytes using autoMACS (Milteyni Biotec, Inc., Auburn, CA). 3 × 106 bone marrow cells after depletion were injected intraperitoneally into 2-day old B6 Rag −/− mice (The Jackson Laboratory, Bar Harbor, ME). After 10 weeks, the inguinal lymph nodes and spleens of these mice were analyzed by flow cytometry, and skin analyzed by histology. For flow cytometry, 3 × 106 cells were blocked using a cocktail of anti-CD16/32 (2.4G2) and normal mouse sera (Jackson ImmunoResearch, West Grove, PA) before being stained with fluorescently labeled antibodies to determine their phenotype and activation status. The following antibody conjugates were used: CD11b PE-TR (Life Technologies Corp.), CD4 V500 (Becton Dickinson, San Jose, CA), CD11c FITC, Gr-1 PECy7, CD3 Alexa Fluor 700, CD8 efluor605, B220 PerCpCy5.5, CD44 efluor450, CD62L APC, CD69 PE (eBioscience). Cells were then washed and re-suspended in 2% FBS in PBS for analysis with flow cytometers (LSR-II and Fortessa, Becton Dickinson).
Leiva A.G., Chen A.L., Devarajan P., Chen Z., Damanpour S., Hall J.A., Bianco A.C., Li J., Badiavas E.V., Zaias J., Miteva M., Romanelli P., Nouri K, & Wikramanayake T.C. (2014). Loss of Mpzl3 Function Causes Various Skin Abnormalities and Greatly Reduced Adipose Depots. The Journal of investigative dermatology, 134(7), 1817-1827.
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2

Measuring Antigen-Specific T Cell Cytokines

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The preparation of single-cell spleens was carried out as previously described [25 (link)]. 7 days after the boost immunization, splenocytes were stimulated with S. Typhimurium LPS (50 ng/µL) for 8 h at 37 °C with 5% CO2 in the presence of a protein transport inhibitor mixture (Thermo Fisher). Cytokine expression of CD4+ T cells was measured using rat anti-mouse CD3-Alexa Fluor 700, CD4-FITC, IFNγ-APC/Cy7, and IL-4-Alexa Fluor 647 (eBioscience). The ratio of cytokine-positive cells in the unstimulated sample was subtracted from the antigen-stimulated value of the same mouse to calculate the percentage of antigen-specific cells.
Han Y., Luo P., Zeng H., Wang P., Xu J., Chen P., Chen X., Chen Y., Cao Q., Zhai R., Xia J., Deng S., Cheng A., Cheng C, & Song H. (2023). The effect of O-antigen length determinant wzz on the immunogenicity of Salmonella Typhimurium for Escherichia coli O2 O-polysaccharides delivery. Veterinary Research, 54, 15.
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3

Multiparameter Flow Cytometry of Lymph Node T-cells

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Mediastinal lymph nodes were collected and perfused with dissociation enzyme mix (Miltenyi Biotech, Auburn, CA), finely chopped and incubated for 20 minutes at 37 °C. After enzymatic digestion and red blood cell lysis, samples were incubated with an antibody cocktail consisting of: CD4 FITC, CD3 Alexa Fluor 700, CD25 APC/Cy7 (eBioscience, San Diego, CA) and CD8a BUV 737 (BD Biosciences, San Jose, CA). Cells were then permeabilized and incubated with another antibody cocktail consisting of intracellular markers Foxp3 PE, RORγt PerCP/Cy5.5, T-bet Alexa Fluor 647 (BD Biosciences, San Jose, CA) and GATA-3 PE/Cy7 (R&D Systems, Minneapolis, MN). Cells were analyzed by FACS to determine T-cell subsets in the lymph nodes.
Hall S.C, & Agrawal D.K. (2017). Increased TREM-2 expression on the subsets of CD11c+ cells in the lungs and lymph nodes during allergic airway inflammation. Scientific Reports, 7, 11853.
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4

Adoptive Transfer of Mpzl3-Deficient Lymphocytes

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Bone marrow was flushed out of femurs and tibias of Mpzl3 −/− or +/+ littermates (6 weeks old). The bone marrow was labeled using biotinylated antibodies against CD3, CD4, CD8 and B220 (eBioscience, San Diego, CA). Streptavidin-labeled microbeads were then used to magnetically deplete the biotinylated lymphocytes using autoMACS (Milteyni Biotec, Inc., Auburn, CA). 3 × 106 bone marrow cells after depletion were injected intraperitoneally into 2-day old B6 Rag −/− mice (The Jackson Laboratory, Bar Harbor, ME). After 10 weeks, the inguinal lymph nodes and spleens of these mice were analyzed by flow cytometry, and skin analyzed by histology. For flow cytometry, 3 × 106 cells were blocked using a cocktail of anti-CD16/32 (2.4G2) and normal mouse sera (Jackson ImmunoResearch, West Grove, PA) before being stained with fluorescently labeled antibodies to determine their phenotype and activation status. The following antibody conjugates were used: CD11b PE-TR (Life Technologies Corp.), CD4 V500 (Becton Dickinson, San Jose, CA), CD11c FITC, Gr-1 PECy7, CD3 Alexa Fluor 700, CD8 efluor605, B220 PerCpCy5.5, CD44 efluor450, CD62L APC, CD69 PE (eBioscience). Cells were then washed and re-suspended in 2% FBS in PBS for analysis with flow cytometers (LSR-II and Fortessa, Becton Dickinson).
Leiva A.G., Chen A.L., Devarajan P., Chen Z., Damanpour S., Hall J.A., Bianco A.C., Li J., Badiavas E.V., Zaias J., Miteva M., Romanelli P., Nouri K, & Wikramanayake T.C. (2014). Loss of Mpzl3 Function Causes Various Skin Abnormalities and Greatly Reduced Adipose Depots. The Journal of investigative dermatology, 134(7), 1817-1827.
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5

Multiparametric Flow Cytometry of Immune Cells

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For cell surface markers, cells were stained in PBS containing 2% fetal bovine serum for 15 minutes, unless mentioned otherwise. For intracellular cytokine staining, cells were stimulated with phorbol 12-myristate acetate (PMA) (50 ng/ mL) and ionomycin (1 lg/mL), or with different antigens, the last 8 hours with 10 lg/mL Brefeldin A and/or 2 lmol/mL monensin, at 378C and 5% CO 2 as mentioned above, and then fixed and stained according to manufacturer's protocols. Antibodies used were as follows: CD3-Alexa Fluor 700, CD4-PerCP/Cy5.5A, IFN-c Alexa Fluor 488, IL-17-PerCP/Cy5.5A, CCR7-APCeFluor780 (eBioscience, San Diego, CA, USA), CD8-V500A (Tonbo, San Diego, CA, USA), TNF-a-Alexa Fluor 700(BD), CD45RA-PECy7, CD45RO-PECF594, FAS-PECF594 (BD, San Diego, CA, USA), FasL-PE (Invitrogen), AnnexinV-APC (Immuno Tools, GmbH, Germany). Cells were acquired and analyzed by BD LSR Fortessa II (BD), using FlowJo (Ashland, OR, USA) three-star or FACSDIVA 6 software (BD Biosciences, San Jose, CA, USA).
Tagirasa R., Parmar S., Barik M.R., Devadas S, & Basu S. (2017). Autoreactive T Cells in Immunopathogenesis of TB-Associated Uveitis. Investigative ophthalmology & visual science, 58(13).
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6

Treg Cell Isolation and Expansion Analysis

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Flow cytometry was used to assess Treg cells following isolation from leukapheresis product and following expansion protocols. Antibodies (anti-human) against the following targets were used in the staining and analysis: CD3 Alexa Fluor 700 (Invitrogen), CD4 V500 (BD Biosciences), CD8 eFluor 450 (Invitrogen), CD25 PerCP-Cy 5.5 (BD Biosciences), FOXP3 Alexa Fluor 488 (Invitrogen). Appropriate isotype controls were set for gating schemes and to establish background parameters. Live/Dead fixable blue dead cell UV stain kit was used to assess viability of the cells. For intracellular FOXP3 staining, cells were fixed and permeabilized using the FOXP3/Transcription Factor Staining Buffer Set (BD Biosciences) prior to staining. Cells were subsequently analyzed using a BD LSRII flow cytometer with BD FACSDIVA software.
Thome A.D., Thonhoff J.R., Zhao W., Faridar A., Wang J., Beers D.R, & Appel S.H. (2022). Extracellular Vesicles Derived From Ex Vivo Expanded Regulatory T Cells Modulate In Vitro and In Vivo Inflammation. Frontiers in Immunology, 13, 875825.
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7

Multiparametric Blood Cell Analysis

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Blood was collected from the inguinal veins at the indicated time points (Fig. 1A) and hematological examination was conducted using an autohematology analyzer (Mindray BC-5000; Mindray, Shenzhen, China). Flow cytometric analysis was performed using a BD LSR Fortessa flow cytometer (BD Biosciences, San Jose, CA, USA) and analyzed using FlowJo v10.7.1 (BD Biosciences), as previously described (8 (link)). To exclude dead cells, blood was first stained with Fixable Viability Stain 575V (BD Biosciences) for 20 min at room temperature. For surface staining, cells were stained with the following antibodies for 30 min at 4°C: CD3 (Alexa Fluor 700; Invitrogen, Waltham, MA, USA), CD20 (APC/Cyanine7; Invitrogen), CD27 (PE/Cyanine7; Invitrogen), IgD (PE; BioLegend Inc., San Diego, CA, USA), IgM (FITC; SouthernBiotech, Birmingham, AL, USA), IgG (V450; BD Bioscience), CD4 (V500; Invitrogen), CD8 (V450; Invitrogen), CD95 (PE/Cyanine5; Invitrogen), and CD28 (ECD; Beckman Coulter, Brea, CA, USA). The cells were washed with permeabilization wash buffer and fixed with 1% paraformaldehyde. Data were acquired using an LSR Fortessa system (BD Biosciences) and analyzed using FlowJo v10.7.1 (BD Biosciences).
Baek S.H., Oh H., Koo B.S., Kim G., Hwang E.H., Jung H., An Y.J., Park J.H, & Hong J.J. (2022). Cynomolgus Macaque Model for COVID-19 Delta Variant. Immune Network, 22(6), e48.
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