The Cytek^® cFluor^® Human B Cell Monitoring Kit identifies B cell subsets in human peripheral blood mononuclear cells and in whole blood collected in EDTA, heparin, or CytoChex^® BCT tubes.  This 13-color cFluor B Cell Monitoring assay distinguishes plasmablasts/plasma cells (CD19⁺ CD20^- CD27⁺ CD38⁺⁺), naïve B cells (CD27^- IgD⁺), IgG class switched memory B cells (CD27⁺ IgG⁺) and classifies unswitched memory B cells (CD27⁺ IgD⁺ and/or IgM⁺). Additionally, the kit identifies the T cell (CD3⁺) subsets of CD4⁺ and CD8⁺ T cells. The granulocyte and monocyte markers, CD15 and CD14, facilitate a cleaner lymphocyte gate.

 

PRODUCT DETAILS

Tested Dilution:  5 μL / test

Application:        Flow Cytometry

Storage:               2-8°C and protected from Light. Do not freeze.

Formulation:      Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 0.2% BSA (Origin USA)

Data Sheets:       TDS SDS

EXAMPLE DATA

 

Figure1: Gating strategy for identifying B cell subsets using the Cytek^® cFluor^® Human B Cell Monitoring Kit. Data were generated from normal whole blood collected in heparin. The gating strategy is based on Gatti et al¹. A clean Lymphocyte population was achieved via sequential gates of time, total cells, singlets, not CD15⁺ granulocyte and not CD14⁺ monocyte gates (plots not shown). Lymphocytes are subsetted into CD3⁺ Lymphocytes and CD19⁺ Lymphocytes. CD3⁺ Lymphocytes are further divided into CD4⁺ and CD8⁺ T cells. From the CD19⁺ Lymphocytes, CD19⁺CD20⁺ B cells are selected to identify naïve (CD27^-IgD⁺), class switched memory (CD27⁺IgD^-), class unswitched memory (CD27⁺IgD⁺), and double negative B cells (CD27^-IgD^-). Additionally, from the CD19⁺CD20⁺CD27⁺ memory B cell population, the combination of IgG, IgM and IgD identifies memory IgG class switched cells (IgM^-IgG⁺, IgD^-IgG⁺) and IgM⁺and IgD⁺unswitched (IgM⁺IgG^-, IgD⁺IgG^-). Furthermore, from total CD19⁺ Lymphocytes, CD20^-CD38⁺⁺ cells can be selected to identify B cell precursors (CD38⁺⁺ CD27^-) and plasma cells (CD38⁺⁺ CD27⁺).

 

 

 RECOMMENDED USAGE: 

• Download the cFluor^® Human B Cell Monitoring (Whole Blood) Sample Protocol
• Download the cFluor^® Human B Cell Monitoring (Whole Blood) Template
• Download the cFluor^® Human B Cell Monitoring (PBMC) Sample Protocol 
• Download the cFluor^® Human B Cell Monitoring (PBMC) Template
• Download the cFluor^® Human B Cell Monitoring Acquisition Protocol
• Download the cFluor^® Human B Cell Monitoring Assay Tags Template
• Download the cFluor^® Human B Cell Monitoring Validation Report

 

REFERENCES

1. Gatti A, Buccisano F, Scupoli MT, Brando B. The ISCCA flow protocol for the monitoring of anti-CD20 therapies in autoimmune disorders. Cytometry B Clin Cytom. 2021 Mar;100(2):194-205. doi: 10.1002/cyto.b.21930. Epub 2020 Jun 29. PMID: 32598578.

 

These products are for research use only. Not intended for use in diagnostic procedures.