Anti cd20 apc

Manufactured by Thermo Fisher Scientific

Sourced in Germany

Anti-CD20-APC is a monoclonal antibody conjugated with the fluorescent dye Allophycocyanin (APC). It is designed to bind to the CD20 antigen, which is expressed on the surface of B cells. This product can be used for the identification and analysis of B cells in flow cytometry applications.

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Close spelling variants of this product

CD20-APC Anti-CD20

3 protocols using anti cd20 apc

EBV Recombinant Virus Infection of Primary B Cells

Cited 2 times

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Recombinant EBV KOs, Revs and wild type have been described previously [54 (link)]. Virus production and B cell isolation were performed as previously described [52 (link),55 (link)]. Primary B cells were isolated from anonymous buffy coat residues (UK Blood Transfusion Service). B cell purity was assessed to be >90% CD20+ using anti-CD20-APC (eBioscience) staining and flow cytometric analysis. Three million primary B cells in 1.5 ml were infected with 0.5 ml of supernatant containing a total of 1–3x10⁵ Raji green units [55 (link)]. RNA from three million uninfected primary B cells was taken on the day of infection and infected cells were incubated in RPMI 1640 (Life Technologies) supplemented with 15% foetal bovine serum, penicillin, and streptomycin at 37°C and 5% CO2. For a period of 30 days, every three days—or five days for the second primary B cell infection with RBPJ binding mutant virus (see below) - 0.5 ml of cells were harvested for RNA extraction and replaced by fresh medium. After this, cells were, where possible, grown into LCL and analysed by immunoblotting.

Kalchschmidt J.S., Gillman A.C., Paschos K., Bazot Q., Kempkes B, & Allday M.J. (2016). EBNA3C Directs Recruitment of RBPJ (CBF1) to Chromatin during the Process of Gene Repression in EBV Infected B Cells. PLoS Pathogens, 12(1), e1005383.

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FACS and PLA Analysis of Mouse and Human B Cells

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For FACS analysis of mouse spleen B cells or transfected TKO cells, following fluorophore- conjugated anti-mouse antibodies were used: Anti-CD45R-PerCP-Cy5.5 (RA3-6B2), Anti-IgM-APC, IgM-PE, IgM-FITC (II/41; all eBioscience, Frankfurt, Germany), Anti-CD20-APC, Anti-CD19-PE-Cy7, anti-IgD-AF647, Anti-IgD-APC, Anti-IgD-PE (11-26c.2a; all eBioscience).
For PLA probes against specific targets, the following unlabelled antibodies were used: IgD (11-26c.2a, SouthernBiotech, Birmingham, AL), IgD (AMS9.1; Santa Cruz Biotechnology, Dallas, TX), IgM (R33.24.12, in house hybridoma culture), IgM (rabbit anti-mouse µHC; Rockland Immunochemicals, Gilbertsville, PA), IgM (1B4B1; SouthernBiotech), Lambda light chain (JC5-1; SouthernBiotech), Kappa light chain (187.1; SouthernBiotech), CD19 (6D5; AbD Serotec, Düsseldorf, Germany) and CD20 (AISB12; eBioscience). Igα (HMK7/A9; abcam, Cambridge, UK), Syk (Syk-01; BioLegend, San Diego, CA).
For PLA probes against human BCR, the following unlabelled antibodies were used: IgD (IA6-2; BioLegend), IgD (IADB6) and IgM (SA-DA4) from Acris Antibodies (Herford, Germany), and IgM (Fc5u) from Genway Biotech (San Diego, CA).

Kläsener K., Maity P.C., Hobeika E., Yang J, & Reth M. (2014). B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. eLife, 3, e02069.

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EBNA3C Knockout and Rev Virus Production for Primary B Cell Isolation

Cited 1 time

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Recombinant EBNA3C KO and Rev viruses, virus production, and primary B cell isolation have been described previously (Anderton et al., 2008 (link); Skalska et al., 2013 (link)). B cell purity was assessed to be >90% CD20⁺ using anti–CD20-APC (eBioscience) staining and flow cytometric analysis. RNA was harvested from uninfected primary B cells. Primary B cells (from anonymous donor buffy coats, purchased from the UK Blood Transfusion Service) were infected with virus-containing supernatant, and infected cells were incubated in RPMI 1640 (Thermo Fisher Scientific) supplemented with 15% fetal bovine serum, penicillin, and streptomycin at 37°C and 5% CO₂. Over a period of 30 d, infected cells were harvested for RNA extraction and replaced by fresh medium every 3 or 5 d.

Kalchschmidt J.S., Bashford-Rogers R., Paschos K., Gillman A.C., Styles C.T., Kellam P, & Allday M.J. (2016). Epstein-Barr virus nuclear protein EBNA3C directly induces expression of AID and somatic mutations in B cells. The Journal of Experimental Medicine, 213(6), 921-928.

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