The Cytek^® Tonbo^® Mouse TBNK/Myeloid/Treg Kit is designed by Cytek scientists to enable researchers to quickly perform basic immunophenotyping and analyze B cell, T cell, NK cell, myeloid and regulatory T cell (Treg) populations in mouse samples. This pre-titered ready-to-use mouse kit contains three sets of reagents that are fully optimized for best performance on the Cytek Aurora^™ and Cytek Northern Lights^™ full spectrum systems.

The Cytek^® Tonbo^® Mouse TBNK/Myeloid/Treg Kit can also be used as a backbone to expand to a broader panel.

• Download Flyer

Products required for the kit but not included:

• Cytek^® Tonbo^® Ghost Dye^™ Violet 540
• Cytek^® Tonbo^® Foxp3 / Transcription Factor Staining Buffer Kit

PRODUCT DETAILS

Tube 1: TBNK Identification Tube

Table 1: Composition of Tube 1: TBNK Identification Tube (P/N 95-K101) of the Cytek^® Tonbo^® Mouse TBNK/Myeloid/Treg Kit (P/N 95-K001). Cytek^® Tonbo^® Ghost Dye^™ Violet 540 (P/N 13-0879) is available to purchase separately.

Tube 2: Myeloid Identification Tube

Table 2: Composition of Tube 2: Myeloid Identification Tube (P/N 95-K201) of the Cytek^® Tonbo^® Mouse TBNK/Myeloid/Treg Kit (P/N 95-K001). Cytek^® Tonbo^® Ghost Dye^™ Violet 540 (P/N 13-0879) is available to purchase separately.

Tube 3: Treg Identification Tube

Table 3: Composition of Tube 3: Treg Identification Tube (P/N 95-K301) of the Cytek^® Tonbo^® Mouse TBNK/Myeloid/Treg Kit (P/N 95-K001). Cytek^® Tonbo^® Ghost Dye^™ Violet 540 (P/N 13-0879) and Cytek^® Tonbo^® Foxp3 / Transcription Factor Staining Buffer Kit (P/N TNB-0607-KIT) are available to purchase separately.

DATA

Tube 1: TBNK Identification Tube

Figure 1: Tube 1 enables the identification of lymphocytes, total T cells, helper T cells, cytotoxic T cells, memory and naïve T cells, B cells, and NK cells. Events are first gated on singlets, red blood cell (RBC) exclusion, and viable CD45⁺ cells. The CD45⁺ cells are then divided into CD49b⁺NK1.1⁺ cells which were further segregated into NK cells and NKT cells by expression of CD3. Within not-NK cells, T cells are characterized by expression of CD3 and separated by TCRß. T cell subsets are further characterized by expression of CD4, CD8, CD44, and CD62L to identify helper, cytotoxic, central memory (CM), effector memory (EM), and naïve T cells. In the CD3^- gate, B cells are quantified as B220⁺CD19⁺.

Tube 2: Myeloid Identification Tube

Figure 2: Tube 2 enables the identification of the myeloid subsets: monocytes, macrophages, and neutrophils, in addition to the dendritic cell subsets: conventional type 1 dendritic cells (cDC1), conventional type 2 dendritic cells (cDC2), and plasmacytoid dendritic cells (pDC). Events are first gated on singlets, RBC exclusion, viable CD45⁺ cells, and FSC vs. SSC. CD3^- cells are gated and then divided to characterize myeloid lineage subsets by expression of CD11b, Ly6C, Ly6G, and F4/80 and DC lineage subsets by expression of CD8, CD11c, B220, Ly6C, and CD4.

Tube 3: Treg Identification Tube

Figure 3: Tube 3 enables the identification of regulatory T cells (Tregs) in addition to characterization of immune checkpoint receptors and T cell activation. Intracellular staining of Foxp3 and CD152 is required. Events are first gated on singlets, RBC exclusion, and viable CD45⁺ cells. The CD45⁺CD3⁺ cells are further gated by expression of CD4 and CD8. Within the CD4⁺ T cells, Tregs are identified by co-expression of CD25 and Foxp3. In the T helper (Th) and CD8⁺ populations, immune checkpoint receptor expression can be determined by gating on CD223 (Lag-3), CD279 (PD-1), and CD152 (CTLA-4) subsets while cell activation can be quantified using CD25 and CD69.

RECOMMENDED USAGE

• Download Sample Preparation Guidelines for Cytek^® Tonbo^® Mouse TBNK/Myeloid/Treg Kit
• Download TBNK (Tube 1) Template
• Download Myeloid (Tube 2) Template
• Download Treg (Tube 3) Template

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