膜杰作
Star Staining
描述（Description）
The Human CD32b (Luc) Jurkat Reporter Cell was engineered to not only express the NFAT response element driving luciferase expressing systems, but also express the receptor human CD32b (Gene ID: 2213), 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 CD32b receptor on the surface of Human CD32b (Luc) 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 CD32b on Human CD32b (Luc) Jurkat Reporter Cell by FACS.
Human CD32b (Luc) Jurkat Reporter Cell or negative control cell were stained with PE-Cy7-labeled anti-human CD32b antibody.
Protocol
Application
ADCP response to anti-human CD20 antibody (RLU).
Anti-human CD20 antibody-induced ADCP activity was evaluated using Human CD32b (Luc) Jurkat Reporter Cell in the presence of Raji cells that express CD20 endogenously. The EC50 of anti-human CD20 antibody was approximately 0.112 μg/mL.
Protocol
ADCP response to anti-human CD20 antibody (FOLD).
Anti-human CD20 antibody-induced ADCP activity was evaluated using Human CD32b (Luc) Jurkat Reporter Cell in the presence of Raji cells that express CD20 endogenously. The max induction fold was approximately 226.
Protocol
Passage Stability
Passage stability analysis by Signaling Bioassay.
The continuously growing Human CD32b (Luc) Jurkat Reporter Cell was stimulated with serial dilutions of anti-human CD20 antibody in the presence of Raji cells that express CD20 endogenously. Anti-human CD20 antibody stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 7-25.
Protocol
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背景（Background）
Fcγ receptors (FcγRs), the largest group of FcRs, bind IgG and comprise several subtypes. Low- affinity FcγRs , including two activating receptors (FcγRIIIa (CD16A) and FcγRIIa (CD32A)) and a inhibitory receptor(FcγRIIb (CD32B)), are important mediators of antibody functions in vivo, including antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and induction of cytokines and chemokines. The inhibitory receptor, FcγRIIb (CD32B), contains an immunoreceptor tyrosine-based inhibition motif that counters cell activation when inhibitory and activating receptors become co-engaged.
License Disclosure
This cell line is provided for research use only. This license does not permit you to share, distribute, sell, sublicense, or otherwise make this cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. The license does not permit modification of this cell line in any way. Inappropriate use or distribution of this cell line will result in revocation of the license. Modifications of this cell line, transfer to another facility, or commercial use of the cell lines may require a separate license and additional fees. 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.
