I‘ve been working on western blots using the Protein Simple Wes machine and have had good success with it.
Although recently I had to move to another lab that has also has a Simple Wes machine, but now I’m not getting any good results at all.
I’m using the same method (sample concentrations, antibody dilutions, etc.) I used for the first machine and I’m also titrating antibodies and samples, but I’m not really getting anywhere for the most part.
My guess is the problem is the age of the secondary antibodies that come with the module, but is it possible that the current machine itself that I’m working with is not as sensitive as the first one I worked with?
Thank you.
Hi Brandon,
I used to work for an instrument company in a different field and my boss used to joke, "they bought two of our instruments, ran the same sample, and had the audacity to expect the same results!" Instruments are sold under the pretense that if they meet their specifications, they will measure identically in every way, but the truth is that some are much better than others. Companies will sometimes stop helping you if it meets their specifications.
I'm trying to figure out a similar issue with two Maurice's. Can you tell me what difference you are seeing between your Wes'? Right now I have baseline instability in one Maurice that was recently serviced and I think it is obscuring small peaks.
This guide may help you troubleshoot what is going on with your instrument: https://cdn.ymaws.com/sites/casss.site-ym.com/resource/resmgr/Scientific_Papers/CASSSCETroubleshootingV2.pdf
I’ve never used a Maurice (is that similar to a SimpleWes?).
What I noticed was that the first one I was using gave great clean signal (the bands showed up really well) but the consistency among the samples was off due to BCA issues.
Now I’m getting the exact opposite results, I’m seeing great consistency/accuracy (per the BCA), but I’m getting virtually no signal from the assays, even when I tried diluting one of the primaries to 1/90.
I still think the problem is the secondary and the BCA results, which I’ll be testing soon. I bought new detection modules also so hopefully that helps if that is in fact the problem.
Thank you for your response. I’ll have a look at the link when I can.
The Wes and Maurice are both CE instruments, but the detection schemes are different.
Please update this if you find the solution. My company wanted the Wes to work for our protein, but the results we got back from the applications scientists at ProteinSimple were terrible. We tried troubleshooting with them, but the quantitation was one to two orders of magnitude low compared to SDS-PAGE western blot.
Good luck!
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