Anti cd23 pe

Manufactured by BioLegend

Anti-CD23 PE is a monoclonal antibody that binds to the CD23 antigen expressed on B cells. It is conjugated with the fluorescent dye phycoerythrin (PE) and can be used to detect and analyze CD23-expressing cells by flow cytometry.

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3 protocols using anti cd23 pe

Multiparametric Flow Cytometry of Immune Cell Subsets

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Single cell suspensions of total splenocytes, enriched B cells or BM were stained with different combinations of the following antibodies: Anti-CD4 APC-eFluor 780, anti-CD8 APC-eFluor 780, anti-Gr1 APC-eFluor 780, anti-F4/80 APC-eFluor 780, anti-B220 APC, anti-B220 APC-eFlour 780, anti-B220 FITC, anti CD19 PeCy7, anti-CD38 Alexa Fluor 700, anti-CD93 APC, anti-IgM PerCP-eFluor 710, anti CD21/CD35 eFluor 450 (eBiosciences), anti-CD23 PE (BioLegend), anti-CD4 PE-CF594, anti-CD8 PE-CF594, anti-Ly-6G and Ly-6C PE-CF594 and anti-IgG1 BV421 (BD biosciences). Live dead aqua stain was added to separate dead cells (Life Technologies) and eOD-GT8-specific cells were visualized by the addition of FITC-conjugated eOD-GT8 and PE-conjugated eOD-GT8 CD4bs knock-out. BG505 SOSIP- (Sok et al., 2014 (link)) and 2cc-specific memory B cells were visualized by the addition of biotinylated protein with the addition of streptavidin conjugated PE and APC respectively (BD Biosicences).

Dosenovic P., von Boehmer L., Escolano A., Jardine J., Freund N.T., Gitlin A.D., McGuire A.T., Kulp D.W., Oliveira T., Scharf L., Pietzsch J., Gray M.D., Cupo A., van Gils M.J., Yao K.H., Liu C., Gazumyan A., Seaman M.S., Björkman P.J., Sanders R.W., Moore J.P., Stamatatos L., Schief W.R, & Nussenzweig M.C. (2015). Immunization for HIV-1 Broadly Neutralizing Antibodies in Human Ig Knock-In Mice. Cell, 161(7), 1505-1515.

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Comprehensive Multicolor Immunophenotyping

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Cells from spleens were stained with cocktails of the following anti-mouse antibodies: anti-CD4 APC-eFluor 780, anti-CD8 APC-eFluor 780, anti-Ly-6G (Gr1) APC-eFluor 780, anti- F4/80 APC-eFluor 780, anti- B220 FITC, anti-CD38 Alexa Fluor 700, anti- IgM PerCP-eFluor 710, anti-CD21/CD35 eFluor 450 and anti-IgG1 BV421 (BD Biosciences); anti-CD23 PE (BioLegend); anti IgD PE/Cy7 and anti Ig light chain λ (Biolegend). Dead cells were excluded by staining with the Live-dead aqua stain (Life Technologies). Stained cell samples were analyzed in a BD LSR Fortessa analyzer.

Escolano A., Steichen J.M., Dosenovic P., Kulp D.W., Golijanin J., Sok D., Freund N.T., Gitlin A.D., Oliveira T., Araki T., Lowe S., Chen S.T., Heinemann J., Yao K.H., Georgeson E., Saye-Francisco K.L., Gazumyan A., Adachi Y., Kubitz M., Burton D.R., Schief W.R, & Nussenzweig M.C. (2016). Sequential Immunization Elicits Broadly Neutralizing anti-HIV-1 Antibodies in Ig Knock-in Mice. Cell, 166(6), 1445-1458.e12.

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Quantifying T and B Cell Populations

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To measure T and B cell numbers, WT and STAT6 KO mice were euthanized, and spleens were harvested. Spleens were mechanically disrupted to obtain a splenocyte suspension. Splenocytes were stained with eBioscience Fixable Viability Dye eFluor 780 (Thermo Fisher Cat No. 65-0865-14) in PBS to exclude dead cells. Splenocytes were washed once and stained with the following antibodies for 30 min: anti-CD4 Alexa Fluor 488 (BioLegend Cat: 100529), anti-CD8a Brilliant Violet 510 (BioLegend Cat: 100751), anti-CD19 Brilliant Violet 421 (BioLegend Cat: 115538), anti-CD21 PE/Cy7 (BioLegend Cat: 123419), and anti-CD23 PE (BioLegend Cat: 101607). Cells were then washed once with FACS buffer and analyzed on Attune NxT flow cytometer. Data were analyzed using FlowJo software.

Prakash A., Medved J., Arneja A., Niebuhr C., Li A.N., Tarrah S., Boscia A.R., Burnett E.D., Singh A., Salazar J.E., Xu W., Santhanakrishnan M., Hendrickson J.E, & Luckey C.J. (2023). Class switching is differentially regulated in RBC alloimmunization and vaccination. Transfusion, 63(4), 826-838.

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