Cytek® cFluor® Human B Cell Monitoring Kit
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Description
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 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 saline, pH 7.2, containing 0.09% sodium azide and 0.2% BSA (BSA Country of Origin USA)
The panel contains the following single vial reagents.
Table 1: Composition of the Cytek® cFluor® Human B Cell Monitoring Kit.
DATA
Figure 1. Gating strategy for identifying B cell subsets using the Cytek® cFluor® Human B Cell Monitoring Kit. Data was 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
Human peripheral mononuclear cells (PBMC) or whole blood collected in EDTA, heparin, or CytoChex® BCT tubes have been tested to validate the performance of this kit. Please refer to the section below to download this kit's staining protocols, data acquisition protocols, and experiment templates.
Please briefly centrifuge the reagent vial before use.
Use appropriate personal protective equipment per the product safety data sheet when using this product.
- Download the Sample Preparation Guidelines for the Cytek® cFluor® Human B Cell Monitoring Kit for
- Download the Acquisition Guidelines for the Cytek® cFluor® Human B Cell Monitoring Kit
- Download the Experiment Template for the Cytek® cFluor® Human B Cell Monitoring Kit for
- Download the Assay Tags for the Cytek® cFluor® Human B Cell Monitoring Kit
- Download the Validation Report for the Cytek® cFluor® Human B Cell Monitoring Kit
REFERENCES
1Gatti 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.
For Research Use Only. Not intended for use in diagnostic procedures.
cFluor® B515, cFluor® B532, cFluor® B548, cFluor® BYG610, cFluor® R668, cFluor® R685, and cFluor® R720 are equivalent to CF®488A, CF®503R, CF®514, RPE-CF®596R, CF®647, CF®660C, and CF®700 respectively, manufactured and provided by Biotium, Inc. under an Agreement between Biotium and Cytek (LICENSEE). The manufacture, use, sale, offer for sale, or import of the product is covered by one or more of the patents or pending applications owned or licensed by Biotium. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser's activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel.
cFluor® BYG610, cFluor® BYG667, cFluor® BYG710, and cFluor® BYG781 are tandem dyes made with R-PE. cFluor® R780 is a tandem dye made with APC. Caution: Tandem dyes may show changes in their emission spectra with prolonged exposure to light or fixatives.