Cd23 apc

Manufactured by BioLegend

CD23-APC is a fluorescently-labeled antibody that binds to the CD23 cell surface protein. CD23 is a low-affinity receptor for IgE and is expressed on various cell types, including B cells and activated monocytes. The APC (allophycocyanin) fluorescent label allows for flow cytometric detection and analysis of CD23-expressing cells.

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Anti-CD23-allophycocyanine (APC) (clone B3B4; (2 citations) APC anti-CD23 (2 citations) APC antihuman CD23 (2 citations)

5 protocols using cd23 apc

Multiparametric Flow Cytometry of Murine Immune Cells

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Murine spleens and lymph nodes were dissected and passed through 70 µm strainers. After pre‐incubation with Zombie™ dye and Fc‐block (BioLegend), cells were stained with antibodies against B220‐FITC, B220‐PE‐Cy7, CD19‐FITC, CD25‐APC, CD69‐PerCP‐Cy5.5, CD95‐PE (all BD Bioscience), CD4‐PE, CD8‐FITC, CD11b‐BV510, CD21‐FITC, CD23‐APC, CD40‐PE, CD80‐PerCP‐Cy5.5, CD86‐PE and MHC‐II‐PacificBlue (all BioLegend). T cells were stained for interferon (IFN)‐γ‐APC (BioLegend) and IL‐17A‐PE (BD Bioscience) after 4‐h incubation with 50 ng/mL PMA, 0.5 μg/mL ionomycin and 1 μL/mL Golgi‐Plug; regulatory T cells were determined by intracellular staining for Foxp3‐PE (BD Bioscience).

Traub J., Traffehn S., Ochs J., Häusser‐Kinzel S., Stephan S., Scannevin R., Brück W., Metz I, & Weber M.S. (2019). Dimethyl fumarate impairs differentiated B cells and fosters central nervous system integrity in treatment of multiple sclerosis. Brain Pathology, 29(5), 640-657.

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Splenic B Cell and Bone Marrow Analysis

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To analyze splenic B cells, total splenocytes were obtained by mashing spleens on 70-µm filters followed by red blood cell lysis (Lonza). To analysis red blood cell content in the bone marrow, total bone marrow cells were flushed out of femora and tibiae using PBS. Single-cell suspensions were stained in the presence of Fc receptor-blocking antibodies (mouse CD16/32, clone 93) using the following antibodies (BioLegend): CD19-FITC (clone 1D3/CD19, 152403), B220-PerCP (clone RA3-6B2, 103233), CD93-PE (clone AA4.1, 136503), CD23-APC (clone B3B4, 101619), CD21-Pacific Blue (clone 7E9, 123413), Ter119-FITC (clone TER-119, 116205) and CD45-APC-Cy7 (clone 30-F11, 103115). Cell viability was measured using Zombie Yellow Fixable Viability kit (423103) or DAPI. Flow cytometry data were acquired on the NovoCyte flow cytometer (Acea Biosciences/Agilent Technologies) using NovoExpress (version 1.3.0) and analyzed using FlowJo (BD).

Takahama M., Patil A., Richey G., Cipurko D., Johnson K., Carbonetto P., Plaster M., Pandey S., Cheronis K., Ueda T., Gruenbaum A., Kawamoto T., Stephens M, & Chevrier N. (2024). A pairwise cytokine code explains the organism-wide response to sepsis. Nature Immunology, 25(2), 226-239.

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Multicolor Flow Cytometry Panel

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Commercial antibodies (clones, fluorophores and sources) are as follows: CD3-FITC (145–2C11), CD62L-PE (Mel-14), CD4-PErCP/Cy5.5 (RM4–5), CD8-PerCP/Cy5.5 (53–6.7), CD8-APC (53–6.7), CD209b-APC (22D1), IgM-FITC (II/41), CD4-PE (GK1.5), CD25-AF488 (PC61.5), Streptavidin-PE, CD169-PE/Cy7 (3D6.112) (eBioscience, San Diego, CA); CD11b-FITC (M1/70), CD49d-FITC (R1–2), TCRβ-FITC (GL3), CD21/35-PE (7E9), CD23-APC (B3B4), Ly6G-PE (RB6–8C5), Ly6C-PerCP/Cy5.5 (HK1.4), CD11c-PE/Cy7 (N418), B220-PerCP/Cy5.5 (RA3–6B2), B220-APC (RA3–6B2), B220-APC/Cy7 (RA3–6B2), CD69-BV421 (H1.2F3),CD23-Biotin (B3B4), F4/80-APC (BM8), Ly6G-BV421 (1A8), CD45-Pacific Blue (30-F11), CD45-BV510 (30-F11), CD43-PE (1B11), CD24-PE/Cy7 (M1/69), IgD-Pacific Blue (11–26c.2a), CD69-PE/Cy7 (H1.2F3), IgD-PerCP/Cy5.5 (11–26c.2a), Ly51-AF647 (6C3), CD3-Pacific Blue (17A2), CD3-PE (17A2), TCRγ/δ-biotin (GL3), Streptavidin-PE/Cy7 (Biolegend, San Diego, CA); Siglec-F-PE (E50–2440) (BD Biosciences, San Jose, CA). Samples were preincubated with 1 μg Fc-block (2.4G2 hybridoma; ATCC).
Cells were acquired either on the BD Biosciences LSR Fortessa or with a BD FACScan flow cytometer with DxP multi-color upgrades by Cytek Development Inc. (Woodland Park, NJ) and analyzed using FlowJo software (FlowJo LLC, Ashland, OR).

Anaya E.P., Lin X., Todd E.M., Szasz T.P, & Morley S.C. (2021). Novel mouse model reveals that serine phosphorylation of L-plastin is essential for effective splenic clearance of pneumococcus. Journal of immunology (Baltimore, Md. : 1950), 206(9), 2135-2145.

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Multiparametric Flow Cytometry of Murine and Human B-Cells

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Single‐cell suspensions were stained with a mixture of 5 to 6 fluorophores including and 7‐aminoactinomycin D (7‐AAD) for viability and acquired on the BD Verse flow cytometer with data analyzed using FlowJo software (BD Biosciences). Human B‐cell subsets were identified using the following antibodies: CD19‐APC, CD27‐BV510, CD38‐APC‐Cy7, CD24‐BV421, and IL‐10‐PE‐Cy7 (all from BioLegend). Mouse B‐cell subsets were identified using the following antibodies: B220‐PE, CD23‐APC, CD21‐PE‐Cy7, IgM‐BV421, CD138‐APC, CD86‐PE‐Cy7, CD69‐APC‐Cy7, and IL‐10‐APC‐Cy7 (all from BioLegend). Intracellular IL‐10 expression was detected as previously reported.³³ Immunophenotype of human and murine B‐cell subsets analyzed are listed in Supplementary Table 1.

Lee W., Wang L., Yen M., Hsu P., Lee Y., Liu K., Lin K., Su Y., Sytwu H, & Yen B.L. (2021). Resident vs nonresident multipotent mesenchymal stromal cell interactions with B lymphocytes result in disparate outcomes. Stem Cells Translational Medicine, 10(5), 711-724.

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Multicolor Imaging of Murine Spleen

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For surface staining, frozen spleen sections (10 μm) were dried, fixed in 4% PFA for 10 min, blocked with PBS 5% normal goat serum (NGS), and then incubated with a combination of the following anti‐mouse antibodies in 1% BSA for 1 h: B220 Pacific Blue (RA3‐6 B2), TCRβ AF488 (H57‐597), CD23 APC (B3B4,), IgM AF488 (RMM‐1), F4/80 APC (BM8), CD169 (Siglec‐1) AF555 (MOMA‐1), (BioLegend). Sections were mounted with ProLong Gold antifade reagent (Thermo Scientific). Images were acquired on a LSM 780 (Zeiss) inverted confocal microscope using a Plan‐Apochromat 20× objective.

Lin Y., Pecetta S., Steichen J.M., Kratochvil S., Melzi E., Arnold J., Dougan S.K., Wu L., Kirsch K.H., Nair U., Schief W.R, & Batista F.D. (2018). One‐step CRISPR/Cas9 method for the rapid generation of human antibody heavy chain knock‐in mice. The EMBO Journal, 37(18), e99243.

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