Anti cd20 apc cy7 clone l27

Manufactured by BD

Anti-CD20-APC-Cy7 (clone L27) is a fluorescently-labeled monoclonal antibody that binds to the CD20 antigen on the surface of B cells. It can be used for the detection and enumeration of B cells in flow cytometry applications.

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Lab products found in correlation

Perm wash, by BD (1 mentions) Anti cd3 alexa fluor 700 clone sp34.2, by BD (1 mentions) Anti cd4 percp clone l200, by BD (1 mentions) Anti cd95 pe cy5 clone dx2, by BD (1 mentions) Anti pd 1 pe cy7, by BioLegend (1 mentions) Clone eh12.2h7, by BioLegend (1 mentions) Anti ki67 fitc b56, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Facscanto 2 flow cytometer, by BD (1 mentions)

2 protocols using anti cd20 apc cy7 clone l27

Multiparameter Flow Cytometry Assay

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Flow cytometry was performed as described previously (6 (link)). Isolated cells were incubated with predetermined optimized concentrations of anti-CD3-Alexa fluor 700 (clone SP34-2, BD Pharmingen), anti-CD8-Pacific Blue or PE-CF594 (clone RPA-T8, BD Pharmingen), anti-CD20-APC-Cy7 (clone L27, BD Pharmingen), anti-CXCR5-phycoerythrin (PE) (Clone MU5UBEE, eBioscience), anti-CD200-APC (clone OX104, eBioscience), anti-CD4-PerCP (clone L200, BD Pharmingen), anti-CD95-PE-Cy5 (clone DX2, BD Pharmingen), anti-CCR7-APC-Cy7 (clone 3D12, Biolegend), anti-PD-1-PE-Cy7 (clone EH12.2H7, Biolegend), anti-ICOS-Pacific Blue (C398.4A, Biolegend) and Live/Dead dye-Alexa430 (Invitrogen). After staining the cell surface, cells were permeabilized and fixed by BD perm wash (BD Pharmingen) and washed twice. Finally, cells were incubated with anti-Ki67-FITC (B56, BD Pharmingen), washed twice and diluted in 1% PFA. Data were acquired on a LSRII flow cytometer (BD Biosciences) and the data analyzed using FlowJo software (version 9.2 Tree Star).

Hong J.J., Amancha P.K., Rogers K.A., Courtney C.L., Havenar-Daughton C., Crotty S., Ansari A.A, & Villinger F. (2014). Early lymphoid responses and germinal center formation correlates with lower viral load set points and better prognosis of SIV infection. Journal of immunology (Baltimore, Md. : 1950), 193(2), 797-806.

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Peripheral Blood Immunophenotyping by Flow Cytometry

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Blood samples (∼1 ml) were periodically collected in tubes containing the anticoagulant potassium EDTA for peripheral blood immunophenotyping by flow cytometry for B lymphocytes (CD45+/CD20+); T lymphocytes (CD45 + CD3+);. Samples were analyzed with a FACSCanto II Flow Cytometer (BD Biosciences) and DIVA 6.0 software. Monoclonal antibodies used for the staining of cells were anti-CD45 (PerCP; clone D058-1283; BD Biosciences), anti-CD20 (APC-Cy7; clone L27; BD Biosciences), and anti-CD3 (FITC; clone SP34; BD Biosciences).
During sample analysis, total lymphocyte populations were identified on the flow cytometer using a heterogeneous lymphocyte gating strategy consisting of CD45 fluorescent staining and side-scatter characteristics (SSC) demarcation (CD45_highSSC_low) to delineate lymphocyte populations. Additionally, a dual-platform methodology was used to enumerate the absolute values for each phenotype. Relative values for each phenotype obtained from the flow cytometer were multiplied by the absolute lymphocyte count from the hematology analysis to enumerate absolute cell counts.

Smith E.J., Olson K., Haber L.J., Varghese B., Duramad P., Tustian A.D., Oyejide A., Kirshner J.R., Canova L., Menon J., Principio J., MacDonald D., Kantrowitz J., Papadopoulos N., Stahl N., Yancopoulos G.D., Thurston G, & Davis S. (2015). A novel, native-format bispecific antibody triggering T-cell killing of B-cells is robustly active in mouse tumor models and cynomolgus monkeys. Scientific Reports, 5, 17943.

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