膜杰作
Star Staining
描述（Description）
The Human TrkA (Luc) HEK293 Reporter Cell was engineered to not only express SRE signaling response element, but also express the receptor full length human TrkA (Gene ID: 4914). When stimulated with human NGF protein, the NGF/TrkA interaction drives SRE-mediated luminescence. Neutralization of biological effect of human NGF protein by corresponding antibody results in a decrease in luminescence.
应用说明（Application）
• Screen for neutralizing antibodies blocking the stimulation of human NGF protein.
• Screen for human TrkA small molecule inhibitor
生长特性（Growth Properties）
Adherent
筛选标记（Selection Marker）
Puromycin (2 μg/mL) + Hygromycin (40 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
Expression analysis of human TrkA on Human TrkA (Luc) HEK293 Reporter Cell by FACS.
Cell surface staining was performed on Human TrkA (Luc) HEK293 Reporter Cell or negative control cell using PE-labeled anti-human TrkA antibody.
Protocol
Application
Inhibition of human NGF protein-induced reporter activity by anti-human NGF neutralizing antibody.
This reporter cell was incubated with serial dilutions of antibodies in the presence of human NGF protein (Cat. No. BEF-H5214) with a final concentration of 0.05 μg/mL. The EC50 of anti-human NGF neutralizing antibody is approximately 0.0212 μg/mL
Protocol
Inhibition of human NGF protein-induced reporter activity by human TrkA small molecule inhibitor.
This reporter cell was incubated with serial dilutions of inhibitors in the presence of human NGF protein (Cat. No. BEF-H5214) with a final concentration of 0.03 μg/mL. The EC50 of human TrkA small molecule inhibitor was approximately 6.725 nM.
Protocol
Signaling Bioassay
Response to human NGF protein (RLU).
The Human TrkA (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human NGF protein (Cat. No. BEF-H5214). The EC50 was approximately 0.02679 μg/mL.
Protocol
Response to human NGF protein (FOLD).
The Human TrkA (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human NGF protein (Cat. No. BEF-H5214). The max induction fold was approximately 12.43.
Protocol
Passage Stability
Passage stability analysis by human NGF protein stimulation.
The continuously growing Human TrkA (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human NGF protein (Cat. No. BEF-H5214) stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 13-22.
Protocol
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背景（Background）
Receptor tyrosine kinase involved in the development and the maturation of the central and peripheral nervous systems through regulation of proliferation, differentiation and survival of sympathetic and nervous neurons. High affinity receptor for NGF which is its primary ligand. Can also bind and be activated by NTF3/neurotrophin-3. However, NTF3 only supports axonal extension through NTRK1 but has no effect on neuron survival. Upon dimeric NGF ligand-binding, undergoes homodimerization, autophosphorylation and activation. Recruits, phosphorylates and/or activates several downstream effectors including SHC1, FRS2, SH2B1, SH2B2 and PLCG1 that regulate distinct overlapping signaling cascades driving cell survival and differentiation. In absence of ligand and activation, may promote cell death, making the survival of neurons dependent on trophic factors.
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.
