Cd20 apc

Manufactured by BD

Sourced in United States, Canada

CD20-APC is a fluorescent-labeled antibody that binds to the CD20 surface antigen on B cells. It is used in flow cytometry applications for the identification and enumeration of B cells.

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Lab products found in correlation

Cd4 percp cy5, by BD (2 mentions) Cd19 percp cy5, by BD (2 mentions) Cd3 apc h7, by BD (2 mentions) Anti glyceraldehyde 3 phosphate dehydrogenase gapdh, by Santa Cruz Biotechnology (1 mentions) Bca protein reagent, by Thermo Fisher Scientific (1 mentions) Recombinant human cxcl12, by Thermo Fisher Scientific (1 mentions) Anti actin, by Santa Cruz Biotechnology (1 mentions) Tcrαβ fitc, by BD (1 mentions) Il 21, by Thermo Fisher Scientific (1 mentions) Anti erk1 2, by Cell Signaling Technology (1 mentions) Cd38 percp cy5, by BD (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Ficoll density gradient centrifugation, by BD (1 mentions) Cd3 pacblue, by BD (1 mentions) Cd8 v500, by BD (1 mentions) Cd25 pe cy7, by BD (1 mentions) Cd28 apc, by BD (1 mentions) Cd11a pe, by BD (1 mentions) Cd45ra fitc, by BD (1 mentions)

Close spelling variants of this product

CD20 APC-H7 (clone 2H7, CD20 APC-Cy7 (L27) CD20 APC-H7 CD20 APC (clone L27) Anti-CD20-APC Anti-CD20-APC-Cy7 (clone L27, CD20 APC-Cy7 Anti-CD20-APC H7

6 protocols using cd20 apc

Immune Cell Activation by CXCL12 and IL-21

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Recombinant human CXCL12 and IL-21 were purchased from Peprotech (Rocky Hill, NJ). Peptidoglycan (PGN) poly (I:C), LPS, and R848 were purchased from Sigma-Aldrich (Poole, Dorset, UK). Human CpG-B DNA was purchased from Hycult Biotech (Uden, Netherlands). Flagellin was provided by Dr. Myoung Ho Jang (Osaka University). RPMI 1640 medium and fetal bovine serum (FBS) were purchased from Gibco BRL (Eggenstein, Germany). Anti-ERK1/2 and anti-human phospho-AKT1/2/3 monoclonal antibodies (Ab) were obtained from Cell Signaling Technology. Anti-ß-actin and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) Abs were obtained from Santa Cruz (Paso Robles, CA). Anti-RGS1 Ab was purchased from Novus Biologicals (Littleton, CO). The anti-CXC chemokine receptors (CXCR) 4-APC, TCRαβ-FITC, CD38-PerCP-Cy5.5, and CD20-APC were purchased from BD Biosciences (Heidelberg, Germany). The BCA protein reagent was purchased from Thermo Scientific (Hudson, NH). Interleukin (IL)-2 and IL-4 were provided by Prof. Jongseon Choe (College of Medicine, Kangwon National University).

Pak H.K., Gil M., Lee Y., Lee H., Lee A.N., Roh J, & Park C.S. (2015). Regulator of G Protein Signaling 1 Suppresses CXCL12-Mediated Migration and AKT Activation in RPMI 8226 Human Plasmacytoma Cells and Plasmablasts. PLoS ONE, 10(4), e0124793.

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Flow Cytometric Immune Profiling in Transplant

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Flow cytometric analysis was performed up to 3 times pre-transplant and serially post-transplant to characterize peripheral blood immune cell phenotypes. Total T cells and T cell subsets were quantified by complete blood cell count and flow cytometry. Fresh PBMCs were isolated by Ficoll density gradient centrifugation (BD Biosciences, Franklin Lakes, NJ). PBMCs were stained with the following mAbs: CD3 PacBlue, CD95 V450, CD3 Alexa 700, CD4 PerCP-Cy5.5, CD8 V500, CD28 PE-Cy7, CD25 PE-Cy7, IFNy PE-Cy7, CD28 APC, TNF APC, VLA-4 APC, CD11a PE, CD45RA FITC, CD40 FITC, CCR7 APC, CD20 APC (all BD Biosciences). PBMCs (1.5×10⁶) were incubated with appropriately titered antibodies for 15min at 20°C and washed twice. Samples were acquired immediately on a BD LSR II multicolor flow cytometer (BD Biosciences), and data were analyzed using FlowJo software (Tree Star, San Carlos, CA). For the stimulation assay, 1.5×10⁶ PBMCs were cultured in RPMI 1640 (Corning cellgro, Manassas, VA) supplemented with 10% fetal bovine serum and stimulated with 10µM phorbol 12-myristate 13-acetate (PMA) and 200nM ionomycin (Sigma-Aldrich, St. Louis, MO), with 1ul/ml GolgiPlug protein transport inhibitor for 5 h, +/− IL-15 (10ng/mL). PBMCs were were processed with BD Cytofix/Cytoperm Plus kit (BD 555028) per the manufactures recommendation prior to data acquisition.

Mathews D., Wakwe W., Kim S., Lowe M., Breeden C., Roberts M., Farris A., Strobert E., Jenkins J., Larsen C., Ford M., Townsend R, & Adams A. (2017). Belatacept Resistant Rejection is Associated with CD28+ Memory CD8 T cells. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 17(9), 2285-2299.

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Multiparametric Phenotyping of B Cells

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PBMCs were resuspended in phosphate-buffered saline (PBS), containing 0.5 % (w/v) BSA and 0.01 % sodium azide. PBMCs were incubated with saturating concentrations of fluorescently labeled conjugated monoclonal antibodies (MoAbs). Analysis of cells was performed using a FACSCanto-II flowcytometer and FlowJo software. The following directly conjugated MoAbs were used for flow cytometry: CD19-PerCP-Cy 5.5, CD19 Alexa-700, CD20-APC, CD20-PerCP-Cy 5.5, CD38 PE-Cy7, CD138 APC, IgG-PE, IgD-PE, and IgA-PE from BD-biosciences (San Jose, USA), CD27-FITC from Sanquin (Amsterdam, the Netherlands), IgM-PE from ITK-diagnostics (Uithoorn, the Netherlands).

aan de Kerk D.J., Jansen M.H., Jolles S., Warnatz K., Seneviratne S.L., ten Berge I.J., van Leeuwen E.M, & Kuijpers T.W. (2016). Phenotypic and Functional Comparison of Class Switch Recombination Deficiencies with a Subgroup of Common Variable Immunodeficiencies. Journal of Clinical Immunology, 36(7), 656-666.

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Multiparametric Flow Cytometry of Immune Cells

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EDTA-anticoagulated whole blood samples were stained within 6 h of collection. Briefly, 100 μl of whole blood was stained with CD8 PerCP (SK1), CD4 FITC (SK3), CD3 APC-H7 (SK7), CD20 APC (2H7), PD-1 BV786 (EH12.1), CCR6 BV711 (11A9), inducible costimulator (ICOS) BV650 (DX29), CD21 BV421 (B-ly4), CD45RA PE-Cy7 (HI100), CD27 PE-CF594 (M-T271) (BD Biosciences, CA), CXCR3 BV510 (G025H7) (Biolegend, San Diego, CA), and CXCR5 PE (MU5UBEE) (eBioscience, San Diego, CA) for 15 min, after which red blood cells were lysed with FACS lysing solution (BD Biosciences), washed and resuspended in FACSflow and acquired on a four laser BD LSRFortessa™ X-20 Special Order Research Product (BD Biosciences, San Jose, CA) within 4 h. CS&T beads and mid-range Rainbow Fluorescent Particles (both BD Biosciences) were run before sample acquisition. Compensation was performed for each experiment using BD™ CompBeads (BD Biosciences). Samples were analyzed using FlowJo software version 9.9.6.

Shalekoff S., Loubser S., Dias B.D., Strehlau R., Shiau S., Wang S., He Y., Abrams E.J., Kuhn L, & Tiemessen C.T. (2021). Normalization of B Cell Subsets but Not T Follicular Helper Phenotypes in Infants With Very Early Antiretroviral Treatment. Frontiers in Pediatrics, 9, 618191.

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Characterization of Cryopreserved CLL PBMCs

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Cryopreserved PBMCs from 8 CLL patients with increased absolute lymphocyte count (ALC) levels (≥20x10 9 cells/L) were further characterized. Cells were thawed and labeled with DAPI and with the following surface antibodies: CD45 (Alexa Fluor 700, eBioscience), CD19 (FITC, BD Biosciences), CD5 (PerCP-Cy5.5, BD Biosciences), CD20 (APC, BD Biosciences), HLA-E (PE, Miltenyi Biotec) and isotype control (PE, eBioscience). For the analysis of HLA-G expression on tumor cells, unlabeled antibodies against HLA-G (clone G233) (Invitrogen) and MYC (clone 9E10, used as negative control) were employed, followed by indirect immunofluorescence staining with PE-Cy7 goat anti-mouse IgG (minimal x-reactivity) (Biolegend). Data acquisition was performed in a LSR Fortessa instrument (BD Biosciences) and analyzed using FlowJo. Mean fluorescence intensity (MFI) values were evaluated as shown in Supplemental Methods and in Supplemental Figure 3.

Puiggros A., Blanco G., Muntasell A., Rodríguez-Rivera M., Nonell L., Altadill M., Puigdecanet E., Arnal M., Calvo X., Gimeno E., Abella E., Abrisqueta P., Bosch F., Yélamos J., Ferrer A., López-Botet M, & Espinet B. (2021). Reduced expansion of CD94/NKG2C⁺ NK cells in chronic lymphocytic leukemia and CLL-like monoclonal B-cell lymphocytosis is not related to increased human cytomegalovirus seronegativity or NKG2C deletions. International journal of laboratory hematology, 43(5).

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Flow Cytometry Analysis of Blood Cells

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Absolute numbers of thrombocytes and ferritin levels were routinely measured according to the established diagnostic guidelines. Absolute numbers of lymphocytes were determined with Multitest 6-color reagents (BD Biosciences, San Jose, Calif) according to the manufacturer's instructions. For additional flow cytometry, PBMCs were resuspended in PBS containing 0.5% (wt/vol) BSA and 0.01% sodium azide and then incubated with saturating concentrations of fluorescently labeled conjugated mAb antibodies. Patient samples were analyzed simultaneously with PBMCs from healthy donors. The following directly conjugated mAbs were used: CD3-APC, CD3-APC-H7, CD4-PE-Cy7, CD4-PerCP-Cy5.5, CD8-PerCP-Cy5.5, CD19-PerCP-Cy5.5, CD20-APC, and CD56-FITC (from BD Biosciences); IgD-PE and gamma-1 isotype (from BD Pharmingen, San Diego, Calif); CD20-APC (from Biolegend San Diego, Calif); CD16-FITC and CD27-FITC (from Sanquin, Amsterdam, The Netherlands); and CD45RA-PE (RD-1) (from Beckman Coulter, Brea, Calif). Analysis of cells was performed using a FACS Calibur or Canto-II flow cytometer (from BD Biosciences) and FlowJo software.

Verhoeven D., Schonenberg-Meinema D., Ebstein F., Papendorf J.J., Baars P.A., van Leeuwen E.M.M., Jansen M.H., Lankester A.C., van der Burg M., Florquin S., Maas S.M., van Koningsbruggen S., Krüger E., van den Berg J.M, & Kuijpers T.W. (2022). Hematopoietic stem cell transplantation in a patient with proteasome-associated autoinflammatory syndrome (PRAAS). The Journal of allergy and clinical immunology, 149(3).

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