ADCP Assay CD32/NFAT Jurkat Cell Line (EGFP/GLuc Dual-Reporter)

Product Data:

Figure 1. ADCP Assay CD16/NFAT Jurkat Cell Line (EGFP/GLuc Dual-Reporter) cells were co-cultured with target Raji cells in the presence of 100 ng/ml of anti-CD20 antibody (BioXCell SKU: SIM0008) or an isotype control antibody for 16 hours followed by flow cytometry analysis or luminescence detection of cell culture medium on the Spectramax iD3. 

Key Features:

• Dual-reporter system:
  • EGFP for fluorescence, enabling single-cell resolution ADCP detection via flow cytometry.
  • Gaussia luciferase (GLuc) for luminescence, allowing high-throughput ADCP measurement in microplate format.
• Endogenous NFAT expression.
• Stable CD32A (FcγRIIa; H131 or R131 allotype) expression, validated by flow cytometry.
• Proprietary enhancer-driven dual-reporter for enhanced sensitivity in ADCP assays.

Applications:

• Screening anti-CD32 monoclonal antibodies.
• Assessing ADCP potency in mAbs.
• Research on Fc receptor-related pathways.
• Measuring ADCP activity via flow cytometry (EGFP) or luminometer (GLuc).

Quality Control: Each cell line undergoes rigorous validation, including CD32A expression and NFAT pathway activation, confirmed via flow cytometry and ADCP assays. The stability of expression is verified for at least 20 passages, and the cells are guaranteed mycoplasma-free.

Shipping and Storage: Cells are shipped on dry ice and should be stored in liquid nitrogen vapor phase upon receipt for optimal viability.

How to Use: Co-culture the Jurkat cells with target cells and the antibody of interest. Measure NFAT activation through luminescence.

Contents:

• 1 vial containing >1 x 10^6 ADCP Assay CD32/NFAT Jurkat Cells (High or Low Affinity).
• 1 ml of Blasticidin (10 mg/ml).

Materials Required but Not Supplied:

• Media for Cell Culture:
  • Thaw Medium: RPMI-1640 with 10% FBS and 1% Penicillin/Streptomycin.
  • Growth Medium: RPMI-1640 with 10% FBS, 10 μg/ml Blasticidin
• Materials for Cellular Assays:
  • Antibodies
  • Target cell lines
  • 96-well tissue culture-treated white clear-bottom assay plate.
  • Promega’s Renilla Luciferase Assay System (for detecting Gaussia luciferase activity).

Background:

ADCP is a crucial immune mechanism where effector cells, such as macrophages, engulf target cells coated with antibodies. In this process, CD32A on the effector cells binds to the Fc region of IgG antibodies attached to target cells, leading to activation of the NFAT signaling pathway. This activation triggers phagocytosis, facilitating the internalization and degradation of the target cells. The CD32A receptor exists in two forms—high-affinity (H131) and low-affinity (R131)—which influence the strength of the ADCP response. This cell line allows for detailed analysis of these interactions, making it an essential tool for therapeutic antibody research and development.

These products are covered by a Limited Use License. For more information, please email info@lipexogen.com

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