膜杰作
Star Staining
描述（Description）
The Human IL-10 R alpha/IL-10 R beta (Luc) HEK293 Reporter Cell was engineered to not only express STAT3 signaling response element, but also express the receptors full length human IL-10 R alpha (Gene ID: 3587) and IL-10 R beta (Gene ID: 3588). When stimulated with human IL-10 protein, the IL-10 R alpha/IL-10 R beta interaction drives STAT3-mediated luminescence. Neutralization of biological effect of human IL-10 protein by corresponding antibody results in a decrease in luminescence.
应用说明（Application）
Screen for neutralizing antibodies blocking the stimulation of human IL-10 protein.
Bioactivity detection of human IL-10 fusion protein
生长特性（Growth Properties）
Adherent
筛选标记（Selection Marker）
Puromycin (2 μg/mL) + Hygromycin (40 μg/mL) + Zeocin (20 μg/mL)
培养基（Culture Medium）
DMEM medium + 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
Co-expression analysis of human IL-10 R alpha and IL-10 R beta on Human IL-10 R alpha/IL-10 R beta (Luc) HEK293 Reporter Cell by FACS.
Cell surface staining was performed on Human IL-10 R alpha/IL-10 R beta (Luc) HEK293 Reporter Cell or negative control cell using PE-labeled anti- IL-10 R alpha antibody and APC-labeled anti- IL-10 R beta antibody.
Protocol
Application
Inhibition of human IL-10 protein-induced reporter activity by anti-human IL-10 neutralizing antibody.
This reporter cell was incubated with serial dilutions of antibodies in the presence of human IL-10 protein (Cat.No.IL0-H5219) with a final concentration of 0.004 μg/mL. The EC50 of anti-human IL-10 neutralizing antibody is approximately 0.4839 μg/mL.
Protocol
Signaling Bioassay
Response to human IL-10 protein (RLU).
The Human IL-10 R alpha/IL-10 R beta (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human IL-10 protein (Cat.No.IL0-H5219). The EC50 was approximately 0.0135 μg/mL.
Protocol
Response to human IL-10 protein (FOLD).
The Human IL-10 R alpha/IL-10 R beta (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human IL-10 protein (Cat.No.IL0-H5219). The max induction fold was approximately 240.
Protocol
Passage Stability
Passage stability analysis by Signaling Bioassay.
The continuously growing Human IL-10 R alpha/IL-10 R beta (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human IL-10 protein. Human IL-10 protein stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 5-20.
Protocol
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背景（Background）
Interleukin-10 receptor subunit alpha (IL-10 R alpha ) is a cell surface receptor for the cytokine IL-10 that participates in IL-10 mediated anti-inflammatory functions, limiting excessive tissue disruption caused by inflammation. Upon binding to IL10, induces a conformational change in IL10RB, allowing IL10RB to bind IL10 as well.In turn, the heterotetrameric assembly complex, composed of two subunits of IL10RA and IL10RB, activates the kinases JAK1 and TYK2 that are constitutively associated with IL10RA and IL10RB respectively.These kinases then phosphorylate specific tyrosine residues in the intracellular domain in IL10RA leading to the recruitment and subsequent phosphorylation of STAT3.
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.
