Anti cd21

Manufactured by Thermo Fisher Scientific

Sourced in United States

Anti-CD21 is a laboratory reagent used for the detection and analysis of CD21 expression on cells. CD21 is a receptor expressed on the surface of B lymphocytes and plays a role in B cell activation and function. Anti-CD21 can be used in flow cytometry, immunohistochemistry, and other applications to identify and study B cells.

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3 protocols using anti cd21

Immunohistochemical Analysis of Lymph Nodes

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Lymph nodes were fixed in 10% neutral-buffered formalin and embedded in paraffin. Tissue sections were typically 4-μm thick and were examined after staining with hematoxylin and eosin (H&E). For immunohistochemistry, the sections were first boiled in a citrate-based solution to retrieve antigens and subsequently quenched in 3% hydrogen peroxide. The antibodies used included anti-FOXP3 (Cat. MAB8214, R&D systems, Minneapolis, MN), anti-PD-1 (Cat. AF1021, R&D systems), anti-CD3 (Cat. ab5690, Abcam, Cambridge, MA), anti-PAX5 (Cat. sc-1974, Santa Cruz Biotechnology, Dallas, TX), anti-CD4 (Cat. 14–0042, eBioscience, San Diego, CA), anti-CD8 (Cat. 14–0081, eBioscience), anti-CD21 (Cat. Ab75985, Abcam), MECA79 (Cat. 53–6036-80, eBioscience) and anti-HA (Cat. 3724, Cell Signaling Technology) antibodies. After biotinylated secondary antibody treatment, visualization was carried out with Vectastain ABC-HRP kit (Vector Laboratories, Burlingame, CA) or ABC-AP kit (Vector Laboratories) in combination with DAB or with VectorRed (Vector Laboratories) respectively.

Lee G.J., Jun Y., Jeon Y.K., Lee D., Lee S, & Kim J. (2022). Mice transgenic for human CTLA4-CD28 fusion gene show proliferation and transformation of ATLL-like and AITL-like T cells. Oncoimmunology, 11(1), 2015170.

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Intracellular Cytokine Staining of Immune Cells

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For intracellular-cytokine staining, single-cell suspended splenocytes were incubated with brefeldin A (eBioscience), followed by an additional 5 h of incubation at 37°C. After surface staining with anti-CD3, anti-CD8, anti-CD11b, anti-CD11c, anti-CD19, anti-CD115, anti-F4/80, anti-Ly6C, anti-Ly6G, anti-MHC-II, and anti-NK1.1 antibodies (all from eBioscience), the cells were fixed with 2% formalin, permeabilized with 0.1% saponin, and stained with anti-TNF antibodies (eBioscience) for 30 min at 4°C. B-cell subsets were detected in single-cell suspensions of splenocytes with anti-CD5, anti-CD19, anti-CD21, anti-CD23, and anti-IgM antibodies (all from eBioscience). BD Calibrite (BD Biosciences, USA) beads were added to the samples before acquisition with a BD LSRFortessa.

Shinde P.V., Xu H.C., Maney S.K., Kloetgen A., Namineni S., Zhuang Y., Honke N., Shaabani N., Bellora N., Doerrenberg M., Trilling M., Pozdeev V.I., van Rooijen N., Scheu S., Pfeffer K., Crocker P.R., Tanaka M., Duggimpudi S., Knolle P., Heikenwalder M., Ruland J., Mak T.W., Brenner D., Pandyra A.A., Hoell J.I., Borkhardt A., Häussinger D., Lang K.S, & Lang P.A. (2018). Tumor Necrosis Factor-Mediated Survival of CD169+ Cells Promotes Immune Activation during Vesicular Stomatitis Virus Infection. Journal of Virology, 92(3), e01637-17.

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Flow Cytometric Analysis of Spleen Immune Cells

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Spleens were harvested and single cell suspensions prepared using 100 µm filters. Flow cytometry for B and T cells was performed as previously described^{35 (link)}. Anti-CD19, anti-B220, anti-CD21, anti-CD23, anti-IgM, anti-CD3, anti-CD4, anti-CD8, anti-IL-17A, anti-IL-4 and anti-IFN-γ antibodies were obtained from eBioscience (San Diego, CA, USA), anti-IgD, anti- CD95, anti-CD138, anti-IgG1 and anti-IgG2ab antibodies from BD BioSciences, anti-IL-10 antibody was purchased from Biolegend (San Diego, CA, USA) and biotinylated peanut agglutinin (PNA) from Sigma-Aldrich (St Louis, USA).
Samples were measured on a FACS Canto II HTS or a LSR II flow cytometer (BD BioSciences) and analysis was performed using FlowJo v7.6 research software (Tree Star Inc. Ashland, OR).

Corneth O.B., Schaper F., Luk F., Asmawidjaja P.S., Mus A.M., Horst G., Heeringa P., Hendriks R.W., Westra J, & Lubberts E. (2019). Lack of IL-17 Receptor A signaling aggravates lymphoproliferation in C57BL/6 lpr mice. Scientific Reports, 9, 4032.

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