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Cd20 percp

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CD20-PerCP is a fluorescently-labeled antibody that specifically binds to the CD20 antigen, a cell surface marker expressed on B cells. This product is intended for use in flow cytometry and other immunoassays.

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Lab products found in correlation

Cd19 fitc, by BioLegend (3 mentions) Cd21 apc, by BioLegend (3 mentions) Facscanto 2, by BD (2 mentions) Cd10 pe cy7, by BioLegend (2 mentions) Cd27 pe, by BioLegend (2 mentions) Anti human cd3 alexa fluor 700 okt3, by Thermo Fisher Scientific (1 mentions) Fcr blocking reagent, by Miltenyi Biotec (1 mentions) Facscelesta, by BD (1 mentions) Cd14 bv650, by BioLegend (1 mentions) Cd45ra bv605, by BioLegend (1 mentions) Cd38 bv421, by BioLegend (1 mentions) Fixable viability stain 510, by BD (1 mentions) Cd43 apc, by BioLegend (1 mentions) Cd8 percp, by BioLegend (1 mentions) Hla dr bv605, by BioLegend (1 mentions) Cd16 af700, by BioLegend (1 mentions) Cd27 fitc, by BioLegend (1 mentions) Cxcr5 bv421, by BioLegend (1 mentions) Pd1 apc, by BioLegend (1 mentions) Cd4 fitc, by BD (1 mentions)

5 protocols using cd20 percp

1

Expression of α4-integrin on B-cell subsets

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We analyzed the intensity of α4-integrin expression on B-cell subsets using blood samples of HCs. Human peripheral blood mononuclear cells (PBMCs) of 8 HCs (mean age 30.5 years, male:female 5:3) were isolated using Pancoll solution (PAN-Biotech Pancoll human, density: 1.077 g/mL) and stored in liquid nitrogen until further use. After thawing, cells were incubated with the FcR blocking reagent (Miltenyi Biotec, Bergisch Gladbach, Germany) and stained using anti-human CD3-Alexa Fluor 700 (OKT3; eBioscience, San Diego, CA), CD14-Alexa Fluor 700 (M5E2, BioLegend, San Diego, CA), CD20-PerCP (2H7 BioLegend), IgD-PerCP (IA6-2, BioLegend), CD19-APC/Fire 750 (HIB19, BioLegend), CD27-Brilliant Violet 605 (O323; BioLegend), CD38-eFluor 450 (HB7; eBioscience), α4-integrin—(CD49d) FITC (9F10, BioLegend), and FITC Mouse IgG1, κ Isotype Control (MOPC-21, BioLegend). B-Cell subpopulations were analyzed within the CD3−CD14−, CD19+ population. In this article, naive B cells were defined as CD38− CD27−; memory B cells were defined as CD38−CD27+ and plasmablasts as CD38+CD27+. Expression levels of α4-integrin were analyzed as delta-mean fluorescence intensity (ΔMFI) (MFI of VLA-4-FITC-staining − MFI of fluorescence minus one control without α4-integrin—FITC). Flow cytometry (FACS) data were analyzed using FlowJo 10.6.1 software (Ashland, OR).
Schlüter M., Oswald E., Winklmeier S., Meinl I., Havla J., Eichhorn P., Meinl E, & Kümpfel T. (2021). Effects of Natalizumab Therapy on Intrathecal Immunoglobulin G Production Indicate Targeting of Plasmablasts. Neurology® Neuroimmunology & Neuroinflammation, 8(5), e1030.
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2

Multiparametric Immune Profiling of PBMC

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Cryopreserved PBMCs were thawed and left for 1 h at 37 °C in the CO2 incubator. Subsequently the cells were collected and stained with BD Horizon™ Fixable Viability Stain 510 to identify live cells, as per manufacture protocol. The cells were then washed, and surface stained for IL-21R PE expression on, (i) CD4, CD8 and Tfh (CD4/CD45RA -/CXCR5+/PD1+); (ii) B cell CD20, B1(CD20+CD27+CD43+), Plasma blast (CD20+CD38+) and iii) monocytes CD14+HLADR+.
After staining, the cells were washed and fixed using 2% PFA. Acquisition was done on BD FACSCelesta (Becton-Dickenson, San Jose, CA). Forward and side scatters and singlets were used to gate and exclude cellular debris. The flow cytometry results were analyzed using FlowJo™ v10.8 Software (BD Life Sciences, Ashland, OR). The details of the antibodies used are as follows: Fixable Viability stain510, CD4 FITC (clone: RPA-T4) from BD Bioscience (San Jose, CA), IL-21R PE (clone: 2G1-K12), CD8 PerCP (clone: SK1), PD1 APC (clone: EH12.2H7), CXCR5 BV421 (clone: J252D4), CD45RA BV605 (clone: HI100), CD38 BV421 (clone: HB-7), CD16 AF700 (clone: B73.1), CD14 BV650 (clone: M5E2), HLADR BV605 (clone: L243), CD20 PerCP (clone: 2H7), CD27 FITC (clone: M-T271), CD43 APC (clone: CD43-10G7) from BioLegend (San Diego, CA).
Agrawal S., Baulch J.E., Madan S., Salah S., Cheeks S.N., Krattli RP J.r., Subramanian V.S., Acharya M.M, & Agrawal A. (2022). Impact of IL-21-associated peripheral and brain crosstalk on the Alzheimer’s disease neuropathology. Cellular and Molecular Life Sciences, 79(6), 331.
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3

Characterization of Memory B Cell Subsets

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Fresh PBMCs from subjects in the active TB, LTBI, and eHC groups were used to characterize the subsets of MBCs with a panel of monoclonal antibodies conjugated with fluorochromes. Approximately 1 × 10 6 cells/ml of PBMCs were stained with a cocktail of CD19-FITC, CD20-Per CP, CD21-APC, and CD27-APC/fire 700 (Biolegend, San Diego, CA, USA). The B cells were phenotyped by gating as CD19 + CD20 + CD21 + CD27 + activated MBCs, CD19 + CD20 + CD21 -CD27 + classical MBCs, CD19 + CD20 + CD21 -CD27 - atypical MBCs, and CD19 + CD20 + CD21 + CD27 -naïve B cells. CD19 + CD20 -CD21 + CD27 + plasmablast were excluded in the final analysis ( Portugal et al., 2015 ; (link)Weiss et al., 2009 ) (link). Switched IgG and IgA MBCs were identified using CD19-FITC, CD20-Alexafluro 700, CD21-APC, CD27-APC/fire 700, IgG Per CP/Cy 5.5 (cloneM1310G05) (Biolegend, San Diego, CA, USA), and IgA PE (cloneB3506B4) (SouthernBiotech, Birmingham, USA). The cells were acquired by flow cytometry, FACSCanto II (BD Bioscience, Becton-Dickinson, Oxford, UK), and analyzed using FlowJo software, version 10.5.3 (Tree Star, Ashland, OR, USA).
Soe P.T., Hanthamrongwit J., Saelee C., Kyaw S.P., Khaenam P., Warit S., Satproedprai N., Mahasirimongkol S., Yanai H., Chootong P, & Leepiyasakulchai C. (2021). Circulating IgA/IgG memory B cells against Mycobacterium tuberculosis dormancy-associated antigens Rv2659c and Rv3128c in active and latent tuberculosis. International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases, 110.
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4

B Cell Subset Profiling During Malaria

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PBMCs from during and post-P. vivax infection were used for B cell subset phenotyping. All B cell phenotypic analyses were performed using mouse monoclonal antibodies (mAbs) specific for human B cell markers conjugated to fluorophores as follows: FITC-CD19, PerCP-CD20, PE/Cy7-CD10, PE-CD27, and APC-CD21 (Biolegend). Using this strategy as showed in Supplementary Fig. 4, we observed the proportion of naive B cells as the number of CD10−CD19+CD20+CD21+CD27−cells; plasma cells/blasts were of CD19+CD21−CD20− cells; immature B cells were CD19+CD10+ cells; classical MBCs were CD10−CD19+CD20+CD21+CD27+ cells; atypical MBCs were of CD10−CD19+CD20+CD21−CD27− cells; and activated MBCs were CD10−CD19+CD20+CD21−CD27+ cells. The relative proportions of all the B cell subpopulations as analyzed per total CD19+ B cells for each sample group was determined. FACS analyses were performed on a FACSCanto II flow cytometer (BD Biosciences) using FlowJo software (Tree Star).
Changrob S., McHenry A.M., Nyunt M.H., Sattabongkot J., Han E.T., Adams J.H, & Chootong P. (2018). Persistence of Long-lived Memory B Cells specific to Duffy Binding Protein in individuals exposed to Plasmodium vivax. Scientific Reports, 8, 8347.
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5

B Cell Subsets Profiling in Malaria

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PBMCs collected during acute malaria and 18 months post-infection were used for B cell sub-set phenotyping. Fluorochrome-conjugated, mouse anti-human monoclonal antibodies were used to stain 1 million PBMCs/100 μL FACS buffer. A cocktail consisting of the following mouse monoclonal antibodies was used: FITC-CD19, PerCP-CD20, PE/Cy7-CD10, PE-CD27, and APC-CD21 (Biolegend, San Diego, CA, USA). After staining for 15 min, cells were washed with FACS buffer. Finally, cells were suspended in 250 μL FACS buffer. The analyses were done by a flow cytometer (BD FACSCanto II, Becton–Dickinson Immunocytometry Systems, San Jose, CA, USA). Data were processed using FlowJo software (Tree Star, San Carlos, CA, USA).
Kochayoo P., Changrob S., Wangriatisak K., Lee S.K., Chootong P, & Han E.T. (2019). The persistence of naturally acquired antibodies and memory B cells specific to rhoptry proteins of Plasmodium vivax in patients from areas of low malaria transmission. Malaria Journal, 18, 382.
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