I am trying to express a hydrophobic membrane protein in N. benthamiana plant but the expression level is very low (checked with microscopy) so I doubt may be I need to modify the transformation protocol. Currently, I am using "bioprotocol" (http://www.bio-protocol.org/e95) agroinfiltration method. I am not able to detect desired proteins's band on SDS PAGE after isolating total protein from transformed leaves.
Thank you
Hello, this protocol seems like quite good, you should do a microsomal preparation to detect your protein, by ultracentrifugation for exemple. Good luck !
The protocol you mention seems very good. At which timepoint do you collect agroinfiltrated ssample for protein detection? Try 3-5 days post agroinfiltration, not ealier or later. Also, agroinfiltrating in rdr6 knockout/knockdown would greatly enhance your protein expression. Cheers.
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