Cell Culture Surface Coating with Poly(Methyl Methacrylate) (PMMA)
2:15
3T3 Cell Lysate Preparation
3:26
Adsorbed Protein Layer Effect on Toll-Like Receptor (TLR)-Mediated Nuclear Factor-kappa-B (NF-kappa-B) Macrophage Activity Assessment
6:01
Results: Representative Lysate TLR Ligand Adsorption to Different Polymer Surfaces
7:18
Conclusion
Transcript
This protocol enables the rapid assessment of NF-kappa-B and AP-1 activity in reporter macrophages cultured on adsorbed protein layers and the contribution of cell signaling pathways, like Toll-like receptors, to that response. The main advantage
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This protocol provides researchers with a rapid, indirect method of measuring TLR-dependent NF-кB/AP-1 transcription factor activity in a murine macrophage cell line in response to a variety of polymeric surfaces and adsorbed protein layers that model the biomaterial implant microenvironment.