I use 24mm diameter transwell(0.4μm) to culture Caco-2 cells.
I want to detect expression of protein after cells exposed to cytokine.
there are three methods
first is directly put the lysis buffer into the transwell,but I am worried about part of protein will be lost because of permeablity of membrane.and the membrane maybe broken if I scrape too hard
second is use trypsin to digest cells,then collect them in a centrifuge tube and add the lysis buffer;
third is cut the membrane out from transwell then put it in a centrifuge tube and add the lysis buffer.That means the transwell will not be used again.
Which method is best? Or anyother method?
Hi, are you using 35S Met/35S Cys to detect de novo proteins?
I just use western blot to prove the protein involving in cellular signal transduction.
I don't know about the 35S Met/35S Cys,could you tell me more detials?
Hello, do you imply that you’ll use the transwell after culture the cell on it?
I used to cut down the transwell membrane and perform yhe protein lysis steps.
Our company develops 3D cell barrier culture systems for in vitro study of virtually any cell barrier, and the system comes equipped with both a Fluid Perfusion Unit and a Trans-Endothelial/Epithelial Electrical Resistance (TEER) Measurement Unit! The use of a vessel-shaped 3D environment has been proven more effective than 2D systems like Transwell (
Article Santaguida S, Janigro D, Hossain M, Oby E, Rapp E, Cucullo L...
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