BAL cells were stained separately with two antibody cocktails. The first was for NK cells with NK1.1-PE (BD PharMingen), T cells with TCRβ-APC (BD PharMingen) and NKT cells NK1.1-PE and TCRβ-APC. The second stain was for neutrophils with Ly6G-FITC (BD PharMingen) and B cells with B220-PE (Biolegend). NKT cells were identified using CD1dα-GALCER tetramer conjugated to APC. Propidium Iodide was used to exclude dead cells before data were acquired using either FACSCalibur or FACSCantoII.
Tcrβ apc
Manufactured by BD
The TCRβ-APC is a lab equipment product used for the detection and analysis of T cell receptor beta chain (TCRβ) expression. It is a fluorescently labeled antibody that binds to the TCRβ chain, allowing for the identification and quantification of T cells in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Ly6g fitc, by BD (1 mentions)
B220 pe, by BioLegend (1 mentions)
Nk1.1 pe, by BD (1 mentions)
Cd4 alexa fluor 700, by Thermo Fisher Scientific (1 mentions)
Flt3 pe, by BD (1 mentions)
Cd34 alexa fluor 700, by BD (1 mentions)
Sca1 apc, by BD (1 mentions)
Lineage cocktail apc, by BD (1 mentions)
Cd8 efluor 450, by BD (1 mentions)
Cd4 pe, by BD (1 mentions)
Cd44 pe cy7, by BD (1 mentions)
Lsrfortessa flow cytometer, by BD (1 mentions)
Facsdiva, by BD (1 mentions)
Cd24 pe, by BD (1 mentions)
Cd25 apc, by BD (1 mentions)
Cd4 percp cy5, by BD (1 mentions)
Cd11c pe cy7, by BD (1 mentions)
Cd11b apc cy7, by BD (1 mentions)
F4 80 apc, by Thermo Fisher Scientific (1 mentions)
Anti cd16 32, by BD (1 mentions)
4 protocols using tcrβ apc
1
Multiparametric Flow Cytometry of Immune Cells
2
Comprehensive Bone Marrow Flow Cytometry
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Flow cytometry was performed on total bone marrow or thymic single cell suspensions. Cells were stained with fluorophore-conjugated antibodies from eBioscience: CD4-Alexa Fluor 700 (56-0041), CD4-PE (12-0041), CD8-eFluor 450 (48-0081), CD24-PE (12-0241), CD25-APC (17-0251), CD34-Alexa Fluor 700 (56-0341), CD44-PE-Cy7 (25-0441), c-Kit-APC-eFluor 710 (47-1171), Flt3-PE (12-1351), IL-7Rα-eFluor 450 (48-1271), Sca1-APC (17-5981) or BD Pharmingen: Lineage Cocktail-APC (558074), Lineage Cocktail-PerCP-Cy5.5 (561317), and TCRβ-APC (561080). Samples were analyzed on an LSR Fortessa flow cytometer (BD Biosciences) in the Flow Cytometry and Cell Sorting Core at BCM. Data interpretation used FACSDiva (BD Biosciences) and FlowJo (TreeStar Inc.) software.
3
Multiparametric Flow Cytometry Analysis of BAL Leukocytes
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Following the centrifugation of BAL fluid to isolate leukocytes, a total cell count was performed using a Neubauer Haemocytometer and trypan blue to exclude dead cells. Samples were then treated with anti-CD16/32 (BD Pharmingen) and stained at 4 °C for 25 min with the following pre conjugated mAbs against the indicated antigens: MHCII-FITC, NK1.1-FITC, CD4-PerCP Cy 5.5, CD11c-PECy7, B220-PE Texas Red, Ly6G-V450, CD44-V450, TCRβ-APC, CD11b-APCCy7, CD3-APC (all BD Biosciences); 7/4-PE (AbD Serotec), CD8-PECy7 (Invitrogen), F4/80-APC (Invitrogen). Samples were then washed before analysis on a CyAn™ ADP Flow cytometer using Summit software (Beckman Coulter). Cell subsets were defined as the following: eosinophils (F4/80intCD11bintSSChi), 7/4− monocytes (F4/80lowCD11bintSSClow7/4−), 7/4+ monocytes (F4/80lowCD11bintSSClow7/4+), myeloid dendritic cells (DCs) (CD11b+CD11c+MHCII+), CD4+ T cells (CD3+αβTCR+CD4+), CD8+ T cells (CD3+αβTCR+CD8+), NK (NK1.1+αβTCR−CD3−) and NKT (NK1.1+αβTCR+CD3+) cells.
4
Multiparametric Flow Cytometry Analysis
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Antibodies used to detect the following mouse proteins were: CD4-PerCP, -647, -FITC (RM4-5), CD8-biotin, -PerCP, -647 (53-6.7), CD11b-biotin (M1/70), CD16/32 purified (2.4G2), CD19-biotin (1D3), CD25-PerCP, -APC, -PE (3C7), Gr1-biotin (RB6-8C5), CD44-BV421, -PE and -APC (IM7); TCRβ-APC, -PE, and -BV421 (H57-597); CD3e-PerCP, -APC and -PE -biotin (2C11), NK1.1-biotin (PK136), pTα purified (2F5), CD3γε-APC (17A2), anti-mouse pZAP70 -647 (Y319), obtained from BD Pharmingen; F4/80biotin (BM8), CD98-PE (RL388), CD8-BV421 (53-6.7), CD71-PE (R71217), all from eBioscience. Annexin V-PE, 7AAD and the APC-labeled anti-BrdU mAb (3D4) were purchased from BD Pharmingen. The APA1/1 monoclonal antibody, which recognizes a conformational epitope of CD3e, and its use as a probe for the conformational change of the TCR have been described previously (Risueno et al., 2005) (Risueno et al., 2005) .
Where necessary, secondary antibodies (anti-rabbit Alexa647 and anti-mouse Alexa647 from ThermoFisher) or fluorescent probes (Streptavidin-PErcp and -APC from BD Pharmingen, Streptavidin-PE from Invitrogen and Streptavidin -BV421 from Biolegend) were used.
Where necessary, secondary antibodies (anti-rabbit Alexa647 and anti-mouse Alexa647 from ThermoFisher) or fluorescent probes (Streptavidin-PErcp and -APC from BD Pharmingen, Streptavidin-PE from Invitrogen and Streptavidin -BV421 from Biolegend) were used.
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