HT-22 cells were lysed in RIPA buffer with phosphate and protease inhibitors
Protein concentration was measured for each OR
Proteins were prepared with Laemmli sample buffer before loading into 4% Bis-Tris gels to measure protein purity and size
Relevant documents
a data-cke-saved-href="https://2017.igem.org/File:Western_Blotting_Protocol.docx" href="https://2017.igem.org/File:Western_Blotting_Protocol.docx">Western Blotting Protocol
Confocal Microscopy
HT-22 cells were fixed with 4% paraformaldehyde and immunohistochemistry with antibodies responsible for individual OR binding followed by secondary antibodies for visualization
Action Potential Measurements
Nucleofected HT-22 cells were stimulated with individual odorants
The neurons generated action potentials that were measured via a microelectrode array