To save class time, the vacuum system is ideal, but with the ultra-sensitive IR detection (Odyssey) the blots are NOT pretty with blotches sometimes obscuring the protein bands. We use a dilution of 0.66 uL per 5 mL in 0.5% BSA in PBS-Tween 20, i.e. scaling down from 1 uL / 15 mL. Is that the problem??
I evaluated the SNAP.id system when I used an Odyssey CLx at my previous institution. I was not able to get signal from even actin, much less a lower expressed gene.
Wet transfer or semi-dry transfer is far superior, even though it takes a little bit longer. I ultimately decided cutting corners/time wasn't worth it if I couldn't trust my experiment. Semi-dry only takes 45 minutes, which saves you 15 minutes if you want. I do not recommend the SNAP.id system unless you already have a set of antibodies that you know work with the system from a colleague, collaborator or trustworthy researcher.
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