Hi, I have some questions about the electrophoresis and transfer condition.
In our lab, we used 15% gel to SDS-page and semi-dry transfer
In electrophoresis, i migrated the stacking gel in 80 V (approximately 15-20 minutes) and subsequently 120 V (1 h 30 min - 2 h).
And I used semi dry transfer machine for 12V 2 hour
In my case, LC3II is detected but LC3I is not detected.
I think in our lab, we used 0.45 um pore size PVDF.
As i know, low molecule protein needs to be 0.2 um pore sized PVDF.
Please tell me your setting (transfer or electrophoresis) and
whether used 0.2 um pore sized PVDF or not.
thank you
Junjie Shao
LC3-II tends to be much more sensitive to be detected by immunoblotting than LC3-I. see belowo
Article How to Interpret LC3 Immunoblotting
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