Palatine tonsils were obtained at the Children’s Hospital of Columbia-Presbyterian Medical Center as residual material from three anonymous patients who had undergone elective tonsillectomy due to chronic tonsillitis in compliance with Regulatory Guideline 45 CFR 46.101 (b) (4 (link)) for Exempt Human Research Subjects of the U.S. Department of Health and Human Services and according to protocols approved by the Columbia University Institutional Ethics Committee. Tonsil specimens were placed on ice immediately after surgical removal. Mononuclear cells (MNC) were isolated by disaggregating tissues in RPMI 1640 medium (Gibco) followed by Ficoll-Isopaque (GE Healthcare) density centrifugation (27 (link)). MNC were stained using the following antibodies: anti-CD38-PE (clone HB7, BD Biosciences), anti-IgD-FITC (clone IA6-2, BD Biosciences), anti-CD3-FITC (clone UCHT1, Beckman Coulter), anti-CD184 (CXCR4)-Brilliant Violet 421 (clone 12G5, BioLegend), anti-CD83-APC (clone HB15e, BioLegend). Total GC (CD38+/IgD-/CD3-), DZ (CD38+/IgD-/CD3-/CXCR4high/CD83low) and LZ (CD38+/IgD-/CD3-/CXCR4low/CD83high) B cells were sorted using an Influx cell sorter (BD Biosciences). Data rendering was performed using FlowJo (TreeStar).
Anti cd38 pe clone hb7
Manufactured by BD
Sourced in United States
Anti-CD38-PE (clone HB7) is a fluorescently-labeled monoclonal antibody that binds to the CD38 antigen. CD38 is a cell surface glycoprotein expressed on various cell types, including plasma cells and activated T and B cells. This antibody can be used for the identification and enumeration of CD38-positive cells in flow cytometry applications.
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Lab products found in correlation
Anti cd3 fitc clone ucht1, by Beckman Coulter (2 mentions)
Anti igd fitc clone ia6 2, by BD (2 mentions)
Anti cd184 cxcr4 brilliant violet 421 clone 12g5, by BioLegend (2 mentions)
Anti cd83 apc clone hb15e, by BioLegend (2 mentions)
Ficoll isopaque, by GE Healthcare (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Influx cell sorter, by BD (1 mentions)
Quantibrite, by BD (1 mentions)
Facsaria flow cytometer, by BD (1 mentions)
Melphalan, by Merck Group (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Anti cd27 pecy7 clone o323, by Thermo Fisher Scientific (1 mentions)
Cell culture media, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc, by Immunostep (1 mentions)
May gr nwald and giemsa, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Draq5, by Biostatus (1 mentions)
4 protocols using anti cd38 pe clone hb7
1
Tonsil B Cell Subsets Isolation
2
Quantifying Cell-Surface CD38 Molecules
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The phycoerythrin (PE) fluorescence quantitation kit Quantibrite™ with anti-CD38-PE (clone HB7), both from BD, were used to estimate the number of cell-surface CD38 molecules by flow cytometry. For daratumumab binding studies, we incubated the cells with 2.5 µg/mL daratumumab or human IgG1 kappa isotype control, and then stained with mouse anti-human IgG Fc or control and analyzed them by flow cytometry.
3
Tonsil Mononuclear Cell Isolation and Flow Cytometry
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Palatine tonsils were obtained at the Children’s Hospital of Columbia–Presbyterian Medical Center as residual material from anonymous patients who had undergone elective tonsillectomy due to chronic tonsillitis in compliance with Regulatory Guideline 45 CFR 46.101 (b)(4) for Exempt Human Research Subjects of the US Department of Health and Human Services and according to protocols approved by the Columbia University Institutional Ethics Committee. Tonsil specimens were placed on ice immediately after surgical removal. Mononuclear cells were isolated from three donors by disaggregating tissues in RPMI 1640 medium (Gibco), followed by Ficoll-Isopaque (GE Healthcare) density centrifugation (Klein et al., 2003 (link)). Tonsillar mononucleated cells were stained using the following antibodies: anti-CD38-PE (clone HB7; BD Biosciences), anti–IgD-FITC (clone IA6-2; BD Biosciences), anti–CD3-FITC (clone UCHT1; Beckman Coulter), anti-CD184 (CXCR4)–Brilliant Violet 421 (clone 12G5; BioLegend), and anti–CD83-APC (clone HB15e; BioLegend). Total GC (CD38+/IgD−/CD3−), DZ (CD38+/IgD−/CD3−/CXCR4high/CD83low), and LZ (CD38+/IgD−/CD3−/CXCR4low/CD83high) B cells were sorted using a FACSAria flow cytometer (BD Biosciences). Data rendering was performed using FlowJo (TreeStar).
4
Immunophenotyping of Multiple Myeloma Cells
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Cell-culture media, serum, and penicillin-streptomycin were purchased from Invitrogen (Carlsbad, CA). Bortezomib was provided by Millennium Pharmaceuticals (Cambridge, MA) and melphalan and doxorubicin were obtained from Sigma-Aldrich (St Louis, MO). Annexin-V–FITC was purchased from Immunostep (Salamanca, Spain). May-Grünwald and Giemsa stains were obtained from Merck (Darmstadt, Germany). The origin of the antibodies used in immunocytochemistry and flow cytometry was as follows: anti-CD138-APC (clone B-B4), used for immunocytochemistry and flow cytometry, from Miltenyi Biotec (Auburn, CA); anti-CD20-FITC (clone L27), anti-CD138-PerCP-Cy5 (clone MI15), anti-CD56-APC (clone NCAM16.2), anti-CD45-AmCyan (clone 2D1) and anti-CD38-PE (clone HB7) from BD Biosciences (San Jose, CA, USA); anti-CD19-PacificBlue (clone HIB19) and anti-CD27-PE-Cy7 (clone O323) antibodies from eBioscience (San Diego CA, USA); anti-CD38-AlexaFluor700 antibody (clone HIT2) from Exbio (Vestec, Czech Republic) and anti-CD138-FITC (clone B-A38) from Cytognos S.L. (Salamanca, Spain). The FITC anti-Ki-67 Set was purchased from BD Biosciences (San Diego, CA) and DRAQ5® was obtained from Biostatus (Leicestershire, UK).
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