B-Cell Research

 

Analysis of B-cell developmental stages in mouse bone marrow
Analysis of B-cell developmental stages in mouse bone marrow

C57BL/6 mouse bone marrow cells were stained with the following fluorescent antibodies: IgM FITC, CD43 APC, BP-1 PE, CD127 PE-Cy™7, CD45R/B220 APC-Cy7, CD24 BD Horizon™ Brilliant Violet™ 421 (BV421), and IgD BD Horizon™ Brilliant Violet™ 510 (BV510), and analyzed using a BD FACSCanto™ II flow cytometer.

Six-color analysis of human peripheral blood cells
Six-color analysis of human peripheral blood cells

Human peripheral blood mononuclear cells (PBMCs) were stained with the following fluorescent antibodies: CD19 PerCP-Cy™5.5, CD20 APC-H7, IgD FITC, CD27 PE-Cy™7, CD38 APC, and CD24 PE, and analyzed using a BD FACSVerse™ flow cytometer.

Ten-color analysis of human peripheral blood cells
Ten-color analysis of human peripheral blood cells

Human PBMCs were stained with the following fluorescent antibodies: CD19 BD Horizon™ Brilliant Violet™ 711, CD20 Alexa Fluor® 700, IgD PE-Cy7, CD27 BV421, CD38 APC, CD24 BD Horizon™ PE-CF594, CD3 BD Horizon™ V500, CD138 PE, IgM BD Horizon™ Brilliant Violet™ 605 (BV605), and IgG FITC, and analyzed using a BD LSRFortessa™ flow cytometer.

Quantitation of immunoglobulins in human serum samples
Quantitation of immunoglobulins in human serum samples

Serum from five donors was tested using BD™ CBA Human Immunoglobulin Flex Sets to analyze total IgG, IgM, and IgA. Data was acquired on a BD FACSArray™ flow cytometer and analyzed using FCAP Array software. Dot plots for a negative control (detector alone) and one of the samples are shown. The results calculated (in g/L) on the basis of standard curves are shown, as well as the expected range.


Phospho-mTOR staining of human cells
Phospho-mTOR staining of human cells

Human peripheral B lymphocytes were stimulated with CpG ODN2395, then fixed with BD Cytofix™ fixation butter, permeabilized using BD Phosflow™ perm buffer III, and stained with anti-mTOR (pS2448) PE or a matching isotype control, and anti-CD20 Alexa Fluor® 647. Cells were analyzed using a BD FACSCanto™ II system.

Bcl-6 expression in mouse lymph node cells
Bcl-6 expression in mouse lymph node cells

Mouse lymph node cells were stained with anti CD45R/B220, CD4, IgD, and CD95, fixed and permeabilized using the BD Pharmingen™ transcription factor buffer set, and stained with anti-Bcl-6 Alexa Fluor® 488. Flow cytometry was performed using a BD™ LSR II system. Bcl-6 expression was observed in germinal center B cells, identified using the CD4B200+IgDloCD95hi phenotype.

Blimp-1 staining in activated mouse splenocytes
Blimp-1 staining in activated mouse splenocytes

B6 mouse splenocytes activated with LPS for 3 days were analyzed using the BD Pharmingen transcription factor buffer set, anti-CD45R/B220 PE, and either anti-Blimp-1 Alexa Fluor® 647 or a matching isotype control. Flow cytometry was performed using a BD FACSCanto II system.

XBP-1s expression in CpG-stimulated human PBMCs
XBP-1s expression in CpG-stimulated human PBMCs

CpG-stimulated human PBMCs were incubated with BD Horizon™ fixable viability stain 450, fixed and permeabilized using the BD Pharmingen transcription factor buffer set, and stained with anti-CD20 PerCP-Cy™5.5, and either anti-XBP-1s PE or a matching isotype control. Flow cytometry was performed using a BD FACSCanto II system.


Five-color B-cell panel: minimal compensation
Five-color B-cell panel: minimal compensation

Human peripheral blood mononuclear cells (PBMCs) were stained with the following fluorescent antibodies: CD19 BUV395, CD27 BV421, IgD FITC, CD38 PE-CF594, and IgM APC. Samples were acquired using a 5-laser BD LSRFortessa™ flow cytometry system.

Analysis of Pax-5 expression in mouse splenic B-cell subpopulations
Analysis of Pax-5 expression in mouse splenic B-cell subpopulations

C57BL/6 mouse splenocytes were stained with the following fluorescent antibodies: CD11b Alexa Fluor® 700, IgM FITC, CD19 BUV395, CD45R/B220 APC, CD93 PE, IgD BD Horizon™ Brilliant Violet™ 605 (BV605), CD21 PerCP Cy™5.5, CD23 BD Horizon™ Brilliant Violet™ 421 (BV421), and CD3 BD Horizon™ Brilliant Violet™ 711 (BV711). Following surface staining, cells were fixed and permeabilized using the BD Pharmingen™ transcription factor buffer set, intracellularly stained with BD Horizon™ Brilliant Violet™ 510 (BV510) anti Pax-5, and analyzed using a special order BD LSRFortessa™ X-20 system.



APC-Cy7: US Patent 5,714,386

Alexa Fluor® is a registered trademark of Life Technologies Corporation.

CF is a trademark of Biotium, Inc.

Cy™ is a trademark of GE Healthcare. Cy™ dyes are subject to proprietary rights of GE Healthcare and Carnegie Mellon University, and are made and sold under license from GE Healthcare only for research and in vitro diagnostic use. Any other use requires a commercial sublicense from GE Healthcare, 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA.