膜杰作
Star Staining
描述（Description）
The Human CD32a (131H) (Luc) Jurkat Reporter Cell was engineered to not only express the NFAT response element driving luciferase expressing systems, but also express the receptor full length human CD32a (131H) (Gene ID: 2212) exhibiting a higher affinity for IgG2 isotypes compared to CD32a-131R, which can use to evaluate ADCP activity of antibodies in the presence of corresponding target cells. When co-cultured with a target cell and relevant antibody, the antibody simultaneously binds the target cell antigen and CD32a (131H) receptor on the surface of Human CD32a (131H) Jurkat Reporter Cell, resulting in receptor clustering, intracellular signaling and NFAT-mediated luminescence.
应用说明（Application）
Determination of ADCP activity induced by antibodies.
生长特性（Growth Properties）
Suspension
筛选标记（Selection Marker）
Puromycin (5 μg/mL) + Hygromycin (20 μg/mL)
培养基（Culture Medium）
RPMI-1640 + 10% FBS
冻存液（Freeze Medium）
Serum-free cell cryopreservation medium
装量（Quantity）
1 vial contains at least 5×10^6 cells in 1 mL serum-free cryopreservation medium
存储（Storage）
Frozen in liquid nitrogen.
支原体检测（Mycoplasma Testing）
Negative
无菌检测（Sterility Testing）
Negative
使用说明（Instructions for Use）
See data sheet for detailed culturing and assay protocol.
Receptor Assay
Expression analysis of human CD32a on Human CD32a (131H) (Luc) Jurkat Reporter Cell by FACS.
Cell surface staining was performed on Human CD32a (131H) (Luc) Jurkat Reporter Cell or negative control cell using PE-labeled anti-human CD32a antibody.
Protocol
Application
ADCP response to anti-human CD20 antibody (RLU).
Anti-human CD20 antibody-induced ADCP activity was evaluated using Human CD32a (131H) (Luc) Jurkat Reporter Cell in the presence of Raji cells that express CD20 endogenously. The EC50 was approximately 0.1054 μg/mL.
Protocol
ADCP response to anti-human CD20 antibody (FOLD).
Anti-human CD20 antibody-induced ADCP activity was evaluated using Human CD32a (131H) (Luc) Jurkat Reporter Cell in the presence of Raji cells that express CD20 endogenously. The max induction fold was approximately 277.90.
Protocol
Passage Stability
Passage stability analysis by Signaling Bioassay.
The continuously growing Human CD32a (131H) (Luc) Jurkat Reporter Cell was stimulated with serial dilutions of Anti-human CD20 antibody in the presence of Raji cells. Anti-human CD20 antibody stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 10-23.
Protocol
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背景（Background）
ADCP is the mechanism by which antibody-opsonized target cells activate the FcγRs on the surface of macrophages to induce phagocytosis, resulting in internalization and degradation of the target cell through phagosome acidification. ADCP is mediated by FcγIIa (CD32a)-activated macrophages leading to augmented phagocytosis.
Permits & Restrictions
This cell line is provided for research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. You are not allowed to share, distribute, sell, modify, sublicense, or otherwise make this cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. AcroBiosystems does not warrant the suitability of this cell line for any particular use, and does not accept any liability in connection with the handling or use of this cell line.
