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, B, 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 new 25-Color Immunoprofiling Assay, Cytek released an 18-color cFluor® reagents kit that can be used with seven reagents from other suppliers. The 25-color assay has been optimized and titrated for use with 7 Brilliant Violet™ reagents available from Biolegend® for analyzing human PBMC and whole blood on Cytek’s Aurora systems equipped with violet, blue, yellow-green, and red lasers.

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.

Data Sheets:  TDS SDS

Validation Report: PDF 

EXAMPLE DATA 

Cell gating for population identification using the Cytek® 25-Color Immunoprofiling Assay Kit. Cells are first gated on singlets, viable cells, CD45⁺, and lymphocytes or monocytes based on scatter. Lymphocytes are divided into T cells, non-T cells, B cells, and ILCs. Monocytes are characterized into phenotypes by expression of CD14 and CD16. Basophils are classified in viable cells.

Within the T lymphocyte population, CD4⁺, CD8⁺, and CD4^-CD8^- T cells can be quantified and further segregated into naïve, memory, effector, and terminal effector subsets.

Additionally, identification and characterization of dendritic cells, NK cells, γδ T cells, NKT-like cells, and regulatory T cells are achievable using the Cytek® 25-Color Immunoprofiling Assay Kit.

Cytek® 25-Color Immunoprofiling Assay, cFluor® Reagent Kit (18C) includes 18 cFluor reagents offered by Cytek

*Immunoprofiling Kit, 7 Color (Brilliant Violet^TM)  includes 7 Brilliant Violet reagents available for purchase from BioLegend

RECOMMENDED USAGE

Human peripheral blood mononuclear cells (PBMC) or whole blood have been tested to validate the performance of this kit. 

• Download sample preparation instructions for PBMCs and Whole Blood
• Download SpectroFlo Experiment Template for PBMCs
• Download SpectroFlo Experiment Template for Whole Blood
• Download SpectroFlo Experiment Template for use with a plate reader
• Download SpectroFlo Experiment Worksheet (NxN plots) for PBMCs
• Download SpectroFlo Experiment Worksheet (NxN plots) for Whole Blood
• Download SpectroFlo Cytek 25-Color Immunoprofiling Assay Tags
• Download SpectroFlo Acquisition Protocol
• Download the 25C IP assay expansion guide
• Download the 25C OMIQ template

REFERENCES

Clevers H et al. Ann Rev Immunol. 1988 6, 629 (1988) 
Evans RL et al. Immunol. 78, 544 (1981) 
Cole DK et al. Cell Mol Immunol. 1, 81 (2004) 
Ferrero E et al. J. Immunol. 145, 331 (1990) 
Wirthmueller U et al. J Exp Med. 175, (1992) 
Dennis O et al. J Immunol. 170, 73 (2003) 
Shevach E. Nat Rev Immunol. 2, 389 (2002) 
Vossen M et al. J Immunol. 180, 3739 (2008) 
Fritsch R et al. J Immunol. 175, 6489 (2005) 
Loken MR et al. Cytometry. 11, 453 (1990) 
Kumar, Connors & Farber. Immunity. 48, 202 (2018) 
Poli A et al. Immunology. 126, 458 (2009) 
Carrette, F & Surh, C. Semin Immunol. 24, 209 (2012) 
Mahnke Y et al. Eur J Immunol. 43, 2797 (2013) 
Goodwin R et al. Cell. 73, 447 (1993) 
Sadler JE. Et al. Thromb Haemost. 78 392-5 (1997) 
Bajzar L, et al. J Biol Chem. 271,16603-8 (1996) 
Walseng E, et al. J Biol Chem. 283,14717 (2008) 
Liu Y, et al. Cells. 9,1206 (2020) 
Kabelitz D, et al. Int Arch Allergy Immunol. 122,1-7 (2000) 
Bonneville M, et al. Nat Rev Immunol. 10,467-478 (2010) 
Sándor, Noémi et al. PloS one vol. 11,9 e0163120 (2016) 
Stacker SA and T.A. Springer. J Immunol. 146, 648 (1991) 
Kared H, et al. Cancer Immunol Immunother. 65, 441-52 (2016) 
Mechtersheimer G, et al. Cancer Res. 51, 1300-7 (1991) 
Stelin, S. et al. J Indian Soc Periodontol 13, 150-4. (2009) 
Macardle PJ, et al. Cell Immunol. 168, 59 (1996) 
Yamada T, et al. J Allergy Clin Immunol. 101, 677 (1998) 
Naeim, F, et al. Atlas of Hematopathology 2nd Edition (2018) 
Esensten, J, et al. Immunity. 44, 973-988 (2016) 
Morandi, F, et al. Cells. 8, 1527 (2019) 
Patsoukis, N, et al. Science Advances. 6, 1-13 (2020) 
Pieper, K, et al. J Allergy Clin Immunol 131, 959-971 (2013) 
Heger, L, et al. Front. Immunol. 11, 559166 (2020) 

For Research Use Only. Not intended for use in diagnostic procedures. 

Cytek dye cFluor® R840 and its conjugates were developed, manufactured, and are commercialized by BioLegend, Inc. under the trademark APC/Fire810™. Cytek dye cFluor® BYG610 was developed, manufactured, and is commercialized by BioLegend, Inc. under the trademark PE-Dazzle 594™. *Fluor conjugated antibody manufactured and supplied by BioLegend Inc.

cFluor® V547, cFluor® B515, cFluor® B532, cFluor® B548, cFluor® YG584, cFluor® R668, cFluor® R685, and cFluor® R720 are equivalent to CF®405L, CF®488A, CF®503R, CF®514, CF®568, 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.

Brilliant Violet™, Brilliant Violet 421™, Brilliant Violet 510™, Brilliant Violet 570™, Brilliant Violet 605™, Brilliant Violet 650™, Brilliant Violet 711™, and Brilliant Violet 785™ are trademarks of Sirigen Group Ltd.

“Cytek,” “SpectroFlo” and “cFluor” are trademarks or registered trademarks of Cytek Biosciences, Inc. All other service marks, trademarks and tradenames appearing herein are the property of their respective owners.