Hi, I'm planning to suggest an update on current western bolt devices, and attracted by the advert of ProteinSimple's new protein assay techque. But i'm confused with multi-target detection on the Wes or Jess system and found little about it.
In tranditonal Western blot, we can perform multiple immunodetection by cutting a whole membrance to bands by protein ladder and incubate with different primary antibodies. So, can i detect different protein of interest, for example EGFR\Akt\actin, in a single run on a SimpleWestern system such as Wes?
Thank you!
Well, I've tried Wes. It can detect different protein in the same capillary, but the effiency strongly depends on antibodies. I tried two antibodies pairs, one worked well (EGFR/GAPDH), another worked really bad. It's a good improvement, but considering the restrict antibody optimazation and consumables economy, I finally chose more tranditional devices
I have been using this equipment for many months now and have come to learn it relatively well.
There is no way to reuse samples after the assay is complete. However, the assay needs very little sample -- 3 ul per well. Because of this, samples can be preserved and the same sample can be tested elsewhere even parallel to testing on the ProteinSimple machine.
Resolution is very good, and it does Total Protein like SDS-PAGE. Some might not want that-- you can't unsee the level of detail. The Jess machine also does replex so you run multiple Westerns restripping for each. As Yang said proper ab selection is key, but that's no different from normal Western.
High throughput and fast, consistent results is an advantage. Avg. less than an hour of sample prep (honestly less than 30m), run takes ~3h. I appreciate the consistency, and quantitative data generated automatically by the system.
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