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Cd19 brilliant violet 421

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The CD19-Brilliant Violet 421 is a fluorescent-labeled antibody that binds to the CD19 cell surface antigen. It is commonly used in flow cytometry applications to identify and enumerate B lymphocytes.

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Lab products found in correlation

Iga fitc, by Bio-Rad (2 mentions) Igm fitc, by Bio-Rad (2 mentions) Cd3 v450, by BD (2 mentions) Igm apc, by BD (2 mentions) Cd20 percp cy5, by BD (2 mentions) Cd38 pe cy7, by BD (2 mentions) Cd19 apc, by BD (2 mentions) Cd27 apc h7, by BD (2 mentions) Dntps, by New England Biolabs (2 mentions) Ribolock, by Thermo Fisher Scientific (2 mentions) Facsaria, by BD (1 mentions) Cd23 pe cy7, by Thermo Fisher Scientific (1 mentions) Igm pe, by Southern Biotech (1 mentions) Ckit cd117 apc, by BioLegend (1 mentions) Cd21 fitc, by BD (1 mentions) B220 fitc, by BD (1 mentions) Cd93 pe cy7, by BioLegend (1 mentions) Cd93 apc, by Thermo Fisher Scientific (1 mentions) Cd25 apc, by BioLegend (1 mentions) Facscanto 2, by BD (1 mentions)

5 protocols using cd19 brilliant violet 421

1

Multicolor Flow Cytometry of Immune Cells

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Bone marrow cells (0.5 × 106) were stained with B220-FITC (Becton Dickinson (BD) Pharmingen, Franklin Lakes, NJ, USA), IgM-PE (Southern Biotechnology Associates, Inc., Birmingham, AL, USA), c-kit (CD117)-APC (Biolegend, San Diego, CA, USA), CD25-APC (Biolegend), CD19-Brilliant Violet 421 (Biolegend), and CD93-PE-Cy7 (Biolegend). Splenocytes (0.5 × 106) were stained with B220-FITC (BD Pharmingen), CD21-FITC (BD Bioscience), IgM-PE (Southern Biotechnology Associates, Inc.), CD93-APC (eBioscience, Vienna, Austria), CD19-Brilliant Violet 421 (Biolegend), and CD23-PE-Cy7 (eBioscience). All cells were analyzed in a FACS Canto II (BD) and data were further processed in Flow Jo version 10.0.4 (Three Star, Inc., Ashland, USA). All analyses started with a singlet gate, thereafter a lymphocyte gate and gates for indicated populations. Results are presented as absolute number of cells of the different populations.
Bernardi A.I., Andersson A., Grahnemo L., Nurkkala-Karlsson M., Ohlsson C., Carlsten H, & Islander U. (2014). Effects of lasofoxifene and bazedoxifene on B cell development and function. Immunity, Inflammation and Disease, 2(4), 214-225.
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2

Isolation of Influenza-specific Plasmablasts

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We collected blood from three individuals (Suppl. Table 1) vaccinated 7 days earlier with the 2010/2011 seasonal trivalent influenza vaccine (Sanofi Pasteur), which consists of 3 strains of inactivated influenza: H1N1 A/California/7/2009, H3N2 A/Perth/16/2009, and B/Brisbane/60/2008. Samples were collected after obtaining informed patient consent and under human subject protocols approved by the Investigational Review Board (IRB) at Stanford University. PBMCs were stained with CD3-V450 (BD 560365), IgA-FITC (AbD Serotec STAR142F or Miltenyi #130-093-071), IgM-FITC (AbD Serotec STAR146F), IgM-APC (BD 551062) or IgM-PE (AbD Serotec STAR146PE), CD20-PerCP-Cy5.5 (BD 340955), CD38-PE-Cy7 (BD 335808), CD19-APC (BD 340437) or CD19-Brilliant Violet 421 (Biolegend 302233), and CD27-APC-H7 (BD 560222). We sorted cells with a BD FACSAria II or III, achieving purities of >80% from the first bulk sort. We gated on CD19+CD20−CD27+CD38++IgA−IgM− cells for the bulk plasmablast sort, and then single-cell sorted them into 96-well PCR plates containing a hypotonic buffer (10mM Tris-HCl pH 7.6) comprised of 2 mM dNTPs (NEB), 5 μM oligo(dT)20VN, and 1 unit/μL of Ribolock (Fermentas), an RNase inhibitor.
Tan Y.C., Scalfone L.K., Kongpachith S., Ju C.H., Cai X., Lindstrom T.M., Sokolove J, & Robinson W.H. (2014). Sequencing Antibody Repertoires Provides Evidence for Original Antigenic Sin Shaping the Antibody Response to Influenza Vaccination. Clinical immunology (Orlando, Fla.), 151(1), 55-65.
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3

Isolation and Culture of Splenic B Cells

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To isolate splenic follicular or marginal zone B cells by FACS, cell suspensions were stained as described for flow cytometric analysis in PBS- 2% FCS. Cells were stained in 15 mL reaction tubes. The solutions’ volumes were adjusted according to the used cell numbers. Cells were stained with CD19 Brilliant Violet 421 (BioLegend), CD23 PE (BioLegend), CD21 biotinylated (eBiosciene) and Streptavidin Cy5 (ImmunoResearch) and isolated with a purity >99% with the MoFlo cell sorter (Beckman Coulter). In addition, splenic naive B cells were isolated by magnetic cell sorting using the “EasySep™ Mouse B Cell Isolation Kit” from STEMCELL according to the manufacturer’s protocol. Isolated splenic B cells were cultured in complete RPMI1640 with 10% FCS and 10 µg/mL LPS with a density of 2x105 cells/mL (37°C, 5% CO2).
Daum P., Ottmann S.R., Meinzinger J., Schulz S.R., Côrte-Real J., Hauke M., Roth E., Schuh W., Mielenz D., Jäck H.M, & Pracht K. (2022). The microRNA processing subunit DGCR8 is required for a T cell-dependent germinal center response. Frontiers in Immunology, 13, 991347.
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4

Multiparametric Flow Cytometry for Immune Profiling

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Cells were stained using fluorochrome-coupled monoclonal antibodies (mAbs). The following mAbs were used in this study. CD3-PECy7, CD3-FITC, CD4-AlexaFluor488, CD4-PerCP/Cy5.5, PD-1-BrilliantViolet421, CD8-APC CD8-PerCP/Cy5.5, CD45RO-PE, CCR7-PE, and CD45RA -AlexaFluor700 were all obtained from BD Bioscience. CD8-FITC, PD-1-PE, PD-1-APC, and CD3-APC were obtained from eBioscience. CD244-PE, CD4-PE/Cy5, CD8-PE/Cy5, CD3-FITC, CD19-FITC, CD19-AlexaFluor700 and CD19-BrilliantViolet421 were obtained from BioLegend. Flow cytometry analyses were performed on a LSRII flow cytometer (BD Biosciences) and subsequently analyzed using Flowjo, version 9.0.2 software (ThreeStar).
Wu J., Xu X., Lee E.J., Shull A.Y., Pei L., Awan F., Wang X., Choi J.H., Deng L., Xin H.B., Zhong W., Liang J., Miao Y., Wu Y., Fan L., Li J., Xu W, & Shi H. (2016). Phenotypic alteration of CD8+ T cells in chronic lymphocytic leukemia is associated with epigenetic reprogramming. Oncotarget, 7(26), 40558-40570.
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5

Isolation and Analysis of IgG+ Plasmablasts from S. aureus Bacteremia Patients

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Blood was collected from individuals with culture-confirmed S. aureus bacteremia after obtaining informed consent and under human subject protocols approved by the Investigational Review Board at Stanford University. For all individuals, blood specimens were obtained within 24 hours after the initiation of standard-of-care antibiotic treatment. PBMCs were isolated using a Ficoll layer and stained with CD3-V450 (BD 560365), IgA-FITC (AbD Serotec STAR142F or Miltenyi #130-093-071), IgM-FITC (AbD Serotec STAR146F), IgM-APC (BD 551062) or IgM-PE (AbD Serotec STAR146PE), CD20-PerCP-Cy5.5 (BD 340955), CD38-PE-Cy7 (BD 335808), CD19-APC (BD 340437) or CD19-Brilliant Violet 421 (Biolegend 302233), and CD27-APC-H7 (BD 560222). IgG+ plasmablasts were gated on CD19+/intCD20-CD27++CD38++IgA-IgM- cells and individually sorted using a BD FACSAria III into a 96-well PCR plate containing a hypotonic lysis buffer (10mM Tris-HCl pH 7.6) containing 2 mM dNTPs (NEB), 5 µM oligo(dT)20VN, and 1 unit/µL of Ribolock (Fermentas), an RNase inhibitor.
Lu D.R., Tan Y.C., Kongpachith S., Cai X., Stein E.A., Lindstrom T.M., Sokolove J, & Robinson W.H. (2014). Identification of Functional Anti-Staphylococcus aureus Antibodies by Sequencing Patient Plasmablast Antibody Repertoires. Clinical immunology (Orlando, Fla.), 152(0), 77-89.
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