Cytek® 25-Color Immunoprofiling Assay, cFluor® Reagent Kit (18C)
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Description
The Cytek® 25-Color Immunoprofiling Assay is designed and optimized by Cytek scientists to provide a turnkey solution for identifying major human immune subpopulations for T cells, B cells, NK cells, monocytes, dendritic cells, and basophils, all of which play important roles in the innate and adaptive immune response in various diseases.
To support the Cytek® 25-Color Immunoprofiling Assay, Cytek released an 18-color cFluor® reagent kit that can be used with seven reagents from other suppliers. The Cytek® 25-Color Immunoprofiling Assay has been optimized and titrated for use with seven Brilliant Violet™ reagents available from BioLegend® for analyzing human PBMC and whole blood on systems equipped with violet, blue, yellow-green, and red lasers.
Products required for the panel but not included:
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.
The panel contains the following single vial reagents.
Table 1: Composition of the Cytek® 25-Color Immunoprofiling Assay, cFluor® Reagent Kit (18C) (P/N R7-40002). Immunoprofiling Kit, 7 Color (Brilliant Violet™) (P/N 900004160) and ViaDye™ Red Fixable Viability Dye Kit (P/N R7-60008) are available to purchase separately.
DATA
Figure 1: Upstream cell gating for population identification. Cells are first gated on A) singlets, B) viable cells, C) CD45+, and D) lymphocytes or monocytes. E) B cells are identified as a subset of CD3-TCRγδ- lymphocytes. G) Innate lymphoid cells and H) B cells and their related populations. F) Basophils are classified within viable cells. I) Monocytes are further segregated into phenotypes by expression of CD14 and CD16.
Figure 2: TH and TC characterization in the Cytek® 25-Color Immunoprofiling Assay. A) Within the T lymphocyte population, CD4+, CD8+, and CD4-CD8- T cells can be quantified. Further characterization of B) CD8+, C) CD4+, and D) CD4-CD8- can segregate naïve, memory, effector, and terminal effector subsets.
Figure 3: Additional characterization of lymphocytic populations. A) Dendritic and C) NK cell populations and subsets can be identified in the CD3-TCRγδ- gate. B) γδ T cells, D) NKT-like cells, and E) regulatory T cells are characterized by expression of their prototypic markers and can be further interrogated for different subsets and activation states.
RECOMMENDED USAGE
Human peripheral mononuclear cells (PBMC), or whole blood, has 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® 25-Color Immunoprofiling Assay for
- Download the Acquisition Protocol for the Cytek® 25-Color Immunoprofiling Assay
- Download the Experiment Template for the Cytek® 25-Color Immunoprofiling Assay for
- Download the Experiment Worksheet (NxN Plots) for the Cytek® 25-Color Immunoprofiling Assay for
- Download the OMIQ Template for the Cytek® 25-Color Immunoprofiling Assay
- Download the Assay Tags for the Cytek® 25-Color Immunoprofiling Assay
- Download the Expansion Guide for the Cytek® 25-Color Immunoprofiling Assay
- Download the Validation Report for the Cytek® 25-Color Immunoprofiling Assay
REFERENCES
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For Research Use Only. Not intended for use in diagnostic procedures.
cFluor® B690 is a tandem dye made with PerCP. cFluor® BYG610, cFluor® BYG667, cFluor® BYG710, and cFluor® BYG781 are tandem dyes made with R-PE. cFluor® R780 and cFluor® R840 are tandem dyes made with APC. Caution – Tandem dyes may show changes in their emission spectra with prolonged exposure to light or fixatives.
cFluor® V547, cFluor® B515, cFluor® B532, cFluor® B548, cFluor® YG584, cFluor® BYG610, cFluor® R668, cFluor® R685, cFluor® R720, and cFluor® R840 are equivalent to CF®405L, CF®488A, CF®503R, CF®514, CF®568, RPE-CF®596R, CF®647, CF®660C, CF®700, and APC-CF®790T 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.
Brilliant Violet™, Brilliant Violet 421™, Brilliant Violet 510™, Brilliant Violet 570™, Brilliant Violet 650™, Brilliant Violet 711™, Brilliant Violet 750™, and Brilliant Violet 785™ are trademarks of Sirigen Group Ltd.