I'm using BEAS-2B as a human bronchial epithelial cell line to test the effect of TGF-beta1. In normal conditions they should be E-cadherin positive but my BEAS-2B aren't. I keep them on MEM medium with 0.5% FCS in a 5%CO2 incubator. I tried them on a coated flask (fibronectin/collagen/BSA) but there I have the same problem.
Some troubleshooting questions, in no particular order:
Dear Bruce,
Thank you for your answer!
I performed STR sequencing and they still have the same characteristics fromas the one from ATCC, so no cross contamination.
I did Western blot and ICC on them and compared with E-cadherin positive HCT8/E11 (colon) and MCF7 (breast), but still no sign...(only my positive control were positive :) )
The last couple of months, I put them on 10% FCS and I have to say my test results were very good, so I better repeat the WB and ICC on new lysate to be sure.
Thank you for all the tips!
Sincerely yours,
Lynn
Didn't realize this was an old thread. Sounds like you have the problem solved.
Hi Lynn,
I am facing a similar problem. Do you think its ok to do TGFb treatment in 10% FCS?
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