Cd23 pe cy7

Manufactured by Thermo Fisher Scientific

Sourced in United States

The CD23-PE-Cy7 is a fluorescent-conjugated monoclonal antibody that binds to the CD23 antigen. CD23 is a low-affinity receptor for IgE expressed on the surface of B cells, activated T cells, and other cell types. The PE-Cy7 fluorescent label allows for the detection and analysis of CD23-positive cells using flow cytometry.

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Close spelling variants of this product

Anti-CD23-PE/Cy7 CD23-PE

5 protocols using cd23 pe cy7

Comprehensive B Cell Immunophenotyping

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Bone marrow of Myce and littermate wild‐type control were harvested into single‐cell suspension with red cell lysis and stained with CD19‐Pacific Blue (1D3, BD, NJ, USA), B220‐PE cy7 (RA3‐6B2, BD), CD24‐APC (M1/69, in house), IgM‐FITC (331.12, in house), CD43‐PE (S7, BD). Spleen red cell lysed single cell suspensions were stained with CD19‐BB700 (1D3, BD), B220‐APC (Thermo Fisher, Waltham, USA), CD23‐PE cy7 (B3B4, Thermo Fisher), CD21‐BV421 (7E9, BioLegend, San Diego, USA), CD138‐PE (281–2, BD). The peritoneal cavity wash was stained with CD19‐PE (ID2, in house), B220‐APC (RA3‐6B2, Thermo Fisher), CD23‐PE cy7 (B3B4, Thermo Fisher), CD5‐BB700 (53–7.3, BD), CD11b‐eFlour450 (M1/70, Thermo Fisher). Lymph node and thymus of single cell suspensions were stained with CD19‐BUV395 (1D3, BD), (TCRβ‐PE (H57‐597, BD), CD4‐BV421 (GK1.5, BioLegend) and CD8a‐PerCp/Cy5.5 (53–6.7, eBioscience, San Diego, USA). Flow cytometry analysis was performed on a BD LSRFortessa X‐20 analyzer (BD).

Chan W.F., Coughlan H.D., Ruhle M., Iannarella N., Alvarado C., Groom J.R., Keenan C.R., Kueh A.J., Wheatley A.K., Smyth G.K., Allan R.S, & Johanson T.M. (2023). Survey of activation‐induced genome architecture reveals a novel enhancer of Myc. Immunology and Cell Biology, 101(4), 345-357.

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Comprehensive Immune Cell Profiling

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The following antibodies were used: CD4-V500, CD11c-PE-Cy7, CD19-V450, CD19-APC-Cy7, CD69-PE-Cy7, IgM-PerCP-Cy5.5, CD45.2-V500, CD45.2-APC (BD Biosciences), IgD-PE, CD45.1-FITC, CD11b-PerCP-Cy5.5, CD21-PB, CD23-PE-Cy7, AA4.1-APC (eBiosciences), and CD86-PB, CD45.1-PB (BioLegend). Following red blood cell lysis, Fc receptor blockade, and staining, cells were processed on a BD FACSVerse or LSR II flow cytometer and analyzed using FlowJo v9.6.4 (Treestar).

Mills R.E., Lam V.C., Tan A., Cresalia N., Oksenberg N., Zikherman J., Anderson M., Weiss A, & Hermiston M.L. (2015). Unbiased modifier screen reveals that signal strength determines the regulatory role murine TLR9 plays in autoantibody production. Journal of immunology (Baltimore, Md. : 1950), 194(8), 3675-3686.

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Multicolor Flow Cytometry of Immune Cells

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Bone marrow cells (0.5 × 10⁶) were stained with B220-FITC (Becton Dickinson (BD) Pharmingen, Franklin Lakes, NJ, USA), IgM-PE (Southern Biotechnology Associates, Inc., Birmingham, AL, USA), c-kit (CD117)-APC (Biolegend, San Diego, CA, USA), CD25-APC (Biolegend), CD19-Brilliant Violet 421 (Biolegend), and CD93-PE-Cy7 (Biolegend). Splenocytes (0.5 × 10⁶) were stained with B220-FITC (BD Pharmingen), CD21-FITC (BD Bioscience), IgM-PE (Southern Biotechnology Associates, Inc.), CD93-APC (eBioscience, Vienna, Austria), CD19-Brilliant Violet 421 (Biolegend), and CD23-PE-Cy7 (eBioscience). All cells were analyzed in a FACS Canto II (BD) and data were further processed in Flow Jo version 10.0.4 (Three Star, Inc., Ashland, USA). All analyses started with a singlet gate, thereafter a lymphocyte gate and gates for indicated populations. Results are presented as absolute number of cells of the different populations.

Bernardi A.I., Andersson A., Grahnemo L., Nurkkala-Karlsson M., Ohlsson C., Carlsten H, & Islander U. (2014). Effects of lasofoxifene and bazedoxifene on B cell development and function. Immunity, Inflammation and Disease, 2(4), 214-225.

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Comprehensive Lymphocyte Phenotyping by Flow Cytometry

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To assess lymphocytes by flow cytometry, single cell suspensions were prepared and incubated with primary antibody for 30 minutes on ice. After staining for surface proteins, cells were incubated with Propidium Iodide (BD Biosciences, San Jose, CA, USA) for 10 minutes as a live/dead stain. After staining, cells were fixed with 0.6% formalin. The antibodies used were CD4-PE, CD5-PE, CD19-FITC, CD69-FITC, CD86-PE, B220-APC, CD93-BB515, CD279-APC, CXCR5-PECy7 (all BD Biosciences), IgM-FITC (Southern Biotech), IgD-APC-Cy7 (BioLegend, San Diego, CA, USA), CD21-eFlour450, and CD23-PE-Cy7 (eBioscience Inc., San Diego, CA, USA). Apoptosis was analyzed with Telford reagent. Flow cytometry was performed with a BD LSRII Flow Cytometer and analyzed with FACSDiva software (BD Biosciences, v.8.0).

Tabor D.E, & Gould K.A. (2016). Estrogen Receptor Alpha Promotes Lupus in (NZB x NZW)F1 Mice in a B cell Intrinsic Manner. Clinical immunology (Orlando, Fla.), 174, 41-52.

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Multiparametric Flow Cytometric Analysis

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Cell suspensions of different organs were lysed with ammonium chloride buffer (0.150 mM NH₄Cl, 0.1 mM EDTA, 0.150 mM KHCO₃) to eliminate erythrocytes followed by staining with CD19-FITC/APC (eBio1D3, 1:100), CD38-PE (clone 90, 1:100), IgM-PE/PECy7 (II/41, 1:200), IgD-eFlour450 (11-26c, 1:30), CD5-APC/Pacific Blue (53-7.3, 1:80), B220-eFlour605 (RA3-6B2, 1:40), CD3-FITC (145-2C11, 1:100), ZAP70-FITC (1E7.2, 1:20), CD21-FITC (eBio8D9, 1:100), CD23-PECy7 (B3B4, 1:160), CD93-PE (AA4.1, 1:80), LIN-Cocktail (CD11b-Biotin (M1/70, 1:130), CD3-Biotin (145-2C11, 1:130), Ter119-Biotin (Ter119, 1:130) and Gr1-Biotin (RB6-8C5, 1:130), Streptavidin-PerCP (1:160), Annexin V-Pacific Blue (1:25) and 7-AAD (1:100) (all from eBioscience). Cells were analysed with the Canto II cytometer (BD Bioscience). Data was obtained with the BD FACSDiva^TM (BD) and analysed with the FlowJo 8.5.3 software. The sequential gating strategy for all FACS panels is provided in Supplementary Fig. 11.

Märklin M., Heitmann J.S., Fuchs A.R., Truckenmüller F.M., Gutknecht M., Bugl S., Saur S.J., Lazarus J., Kohlhofer U., Quintanilla-Martinez L., Rammensee H.G., Salih H.R., Kopp H.G., Haap M., Kirschniak A., Kanz L., Rao A., Wirths S, & Müller M.R. (2017). NFAT2 is a critical regulator of the anergic phenotype in chronic lymphocytic leukaemia. Nature Communications, 8, 755.

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