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Cd3 efluor450 17a2

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CD3 eFluor450 (17A2) is a fluorescent antibody conjugate used for the detection and analysis of T cells in flow cytometry applications. It binds to the CD3 antigen, a key component of the T cell receptor complex, and can be used to identify and quantify T cell populations in biological samples.

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Lab products found in correlation

Facscanto 2, by BD (4 mentions) Fixation and permeabilization buffer, by Thermo Fisher Scientific (2 mentions) Streptavidin pe, by Thermo Fisher Scientific (2 mentions) Apc ifnγ, by Thermo Fisher Scientific (2 mentions) Biotin tcrγδ, by Thermo Fisher Scientific (2 mentions) Cd69 fitc, by Thermo Fisher Scientific (2 mentions) Pecy7 pannk, by Thermo Fisher Scientific (2 mentions) Fixable viability dye, by Thermo Fisher Scientific (2 mentions) Ionomycin, by Merck Group (2 mentions) Golgiplug, by BD (2 mentions) Mhc 2 efluor450, by Thermo Fisher Scientific (2 mentions) Cd24 pe cy7, by Thermo Fisher Scientific (2 mentions) Cd45r efluor450, by Thermo Fisher Scientific (2 mentions) Irf8 percp efluor710, by Thermo Fisher Scientific (2 mentions) Cd135 apc pe cf594, by BD (2 mentions) Cd24 buv496, by BD (2 mentions) Cd127 bv421 sb, by BD (2 mentions) Cd45ra pe 14.8, by BD (2 mentions) Cd317 efluor450 apc, by Thermo Fisher Scientific (2 mentions) Cd172a apc percp efluor710, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

CD3 (17A2, CD3-eFluor450

5 protocols using cd3 efluor450 17a2

1

Lung Single-Cell Immunophenotyping Protocol

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Lung single cell suspensions were prepared using a standardized protocol as previously described38 (link). Cells were stained with the following antibodies from BD Biosciences and eBioscience: eFluor450-CD3 (17A2), PerCP-CD4 (RM4-5), FITC-CD8a (53-6.7), Biotin-TCRγδ (GL3), PE-Streptavidin, APC-IL-17A (eBio17B7), APC-IFNγ (XMG1.2), FITC-CD69 (H1.2F3), PerCP-Cy5.5-Gr1 (RB6-8C5), and PECy7-panNK (DX5). For intracellular staining, cells were stimulated for 5 h with 5 ng/ml PMA and 500 ng/ml ionomycin (Sigma-Aldrich; St. Louis, MO, USA) in the presence of a protein-transport inhibitor (GolgiPlug, BD Biosciences; San Jose, CA, USA). Following stimulation, cells were collected, stained with a fixable viability dye (eBioscience), fixed, permeabilized (Fixation and Permeabilization Buffer; eBioscience), then intracellularly stained. Samples were read using a FACSCanto II (BD Biosciences) flow cytometer and analyzed with FlowJo software (version 7.6.5 for Windows; Tree Star; Ashland, OR, USA). To simultaneously compare both quantity (% of cells) and quality (intensity of staining) of IL-17-producing populations, integrated median fluorescent intensity (iMFI) was calculated by multiplying percent of x cell type (i.e., CD4, CD8, γδ, Gr1, NK) of total IL-17A+ cells multiplied by the IL-17A MFI of x cell type.
Huang H., Saravia J., You D., Shaw A.J, & Cormier S.A. (2014). Impaired gamma delta T cell-derived IL-17A and inflammasome activation during early respiratory syncytial virus infection in infants. Immunology and cell biology, 93(2), 126-135.
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2

Lung Single-Cell Immunophenotyping Protocol

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Lung single cell suspensions were prepared using a standardized protocol as previously described38 (link). Cells were stained with the following antibodies from BD Biosciences and eBioscience: eFluor450-CD3 (17A2), PerCP-CD4 (RM4-5), FITC-CD8a (53-6.7), Biotin-TCRγδ (GL3), PE-Streptavidin, APC-IL-17A (eBio17B7), APC-IFNγ (XMG1.2), FITC-CD69 (H1.2F3), PerCP-Cy5.5-Gr1 (RB6-8C5), and PECy7-panNK (DX5). For intracellular staining, cells were stimulated for 5 h with 5 ng/ml PMA and 500 ng/ml ionomycin (Sigma-Aldrich; St. Louis, MO, USA) in the presence of a protein-transport inhibitor (GolgiPlug, BD Biosciences; San Jose, CA, USA). Following stimulation, cells were collected, stained with a fixable viability dye (eBioscience), fixed, permeabilized (Fixation and Permeabilization Buffer; eBioscience), then intracellularly stained. Samples were read using a FACSCanto II (BD Biosciences) flow cytometer and analyzed with FlowJo software (version 7.6.5 for Windows; Tree Star; Ashland, OR, USA). To simultaneously compare both quantity (% of cells) and quality (intensity of staining) of IL-17-producing populations, integrated median fluorescent intensity (iMFI) was calculated by multiplying percent of x cell type (i.e., CD4, CD8, γδ, Gr1, NK) of total IL-17A+ cells multiplied by the IL-17A MFI of x cell type.
Huang H., Saravia J., You D., Shaw A.J, & Cormier S.A. (2014). Impaired gamma delta T cell-derived IL-17A and inflammasome activation during early respiratory syncytial virus infection in infants. Immunology and cell biology, 93(2), 126-135.
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3

Multiparametric Flow Cytometry Analysis

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Cells were kept at 4°C while staining in PBS with 0.5% BSA and 2 mM EDTA in the presence of CD16/32 block (BD clone 2.4G2).
The following antibodies were purchased from BD: CD45.1 BV711 (A20), CD117 BUV395 (2B8), CD135 APC & PE-CF594 (A2F10.1), MHC II V500, BV421, BV510 (M5/114.15.2), CD24 BUV496 (M1/69), CD127 BV421 (SB/199), CD45RA PE (14.8), CD19 BV42 1(1D3). From eBioscience: CD117 PE-Cy7 (2B8), CD317 eFluor450 & APC (eBio927), CD115 PE (AFS98), APC eFluor780 anti-CD11c (N418), MHC II eFluor450 (M5/114/15/2), CD24 PE-Cy7 (M1/69), CD172a APC & PerCP-eFluor710 (P84), Siglec-H PerCP-eFluor710 (eBio440C), eFluor450 Ter119 (Ter119), CD105 eFluor450 (MJ7/18), Irf8 PerCP-eFluor710 (V3GYWCH), CD45R eFluor450 (RA3-6B2), NK1.1 eFluor450 (PK136), CD3 eFluor450 (17A2), Irf4 PE (3E4). From Tonbo Biosciences: CD45.1 PE-Cy7 (A20), CD115 PE (AFS98), CD11c APC-Cy7 (N418). From BioLegend: CD115 BV711 (ASF98), CD45.1 Alexa Fluor 488 (A20), Flag (DYKDDDDK) APC (L5).
Lineage cell depletion kit was purchased from Miltenyi. Cells were analyzed on FACSCanto II or FACSAria Fusion flow cytometers (BD), and data analyzed with FlowJo software (Tree Star).
Grajales-Reyes G.E., Iwata A., Albring J., Wu X., Tussiwand R., Wumesh K., Kretzer N.M., Briseño C.G., Durai V., Bagadia P., Haldar M., Schönheit J., Rosenbauer F., Murphy T.L, & Murphy K.M. (2015). Batf3 maintains Irf8 autoactivation for commitment of a CD8α+ cDC clonogenic progenitor. Nature immunology, 16(7), 708-717.
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4

Multiparametric Flow Cytometry Analysis

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Cells were kept at 4°C while staining in PBS with 0.5% BSA and 2 mM EDTA in the presence of CD16/32 block (BD clone 2.4G2).
The following antibodies were purchased from BD: CD45.1 BV711 (A20), CD117 BUV395 (2B8), CD135 APC & PE-CF594 (A2F10.1), MHC II V500, BV421, BV510 (M5/114.15.2), CD24 BUV496 (M1/69), CD127 BV421 (SB/199), CD45RA PE (14.8), CD19 BV42 1(1D3). From eBioscience: CD117 PE-Cy7 (2B8), CD317 eFluor450 & APC (eBio927), CD115 PE (AFS98), APC eFluor780 anti-CD11c (N418), MHC II eFluor450 (M5/114/15/2), CD24 PE-Cy7 (M1/69), CD172a APC & PerCP-eFluor710 (P84), Siglec-H PerCP-eFluor710 (eBio440C), eFluor450 Ter119 (Ter119), CD105 eFluor450 (MJ7/18), Irf8 PerCP-eFluor710 (V3GYWCH), CD45R eFluor450 (RA3-6B2), NK1.1 eFluor450 (PK136), CD3 eFluor450 (17A2), Irf4 PE (3E4). From Tonbo Biosciences: CD45.1 PE-Cy7 (A20), CD115 PE (AFS98), CD11c APC-Cy7 (N418). From BioLegend: CD115 BV711 (ASF98), CD45.1 Alexa Fluor 488 (A20), Flag (DYKDDDDK) APC (L5).
Lineage cell depletion kit was purchased from Miltenyi. Cells were analyzed on FACSCanto II or FACSAria Fusion flow cytometers (BD), and data analyzed with FlowJo software (Tree Star).
Grajales-Reyes G.E., Iwata A., Albring J., Wu X., Tussiwand R., Wumesh K., Kretzer N.M., Briseño C.G., Durai V., Bagadia P., Haldar M., Schönheit J., Rosenbauer F., Murphy T.L, & Murphy K.M. (2015). Batf3 maintains Irf8 autoactivation for commitment of a CD8α+ cDC clonogenic progenitor. Nature immunology, 16(7), 708-717.
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5

Comprehensive Flow Cytometry for ASCs

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All flow cytometric analysis was conducted on BD FACS Aria III or LSR II. The data were analyzed using Flowjo software. To stain for ASCs, CD138 APC or PerCP-Cy5.5 (clone 281-2, BD Biosciences or Biolegend), B220 APC-Cy7 (clone RA3-6B2, Biolegend), CD98 PerCP-Cy5.5 or PE (clone 4F2, Biolegend), CD93 APC (clone AA4.1, Biolegend), CD3 Efluor 450 (17A2, eBioscience), and CD184 BV421 or PE (clone 2B11/CXCR4; BD Biosciences) were used. For B-cell lymphoma 2 (BCL-2 PeCy7; clone BCL/10C4; Biolegend) and mantle cell leukemia (MCL-1 APC; clone D2W9E; Cell Signaling Technologies) staining, cytofix/cytoperm kit (BD) was used. CD45.1 PE (clone A20; Invitrogen) and CD45.2 BV450 (clone 104; BD Biosciences) antibodies were used to distinguish donor and recipient cells in chimera studies. Fluorescent counting beads (Spherotech) were used to quantify cell numbers routinely.
Aaron T., Laudermilch E., Benet Z., Ovando L.J., Chandran K, & Fooksman D. (2023). TNF-α Limits Serological Memory by Disrupting the Bone Marrow Niche. Journal of immunology (Baltimore, Md. : 1950), 210(5), 595-608.
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