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ELISAs [Nov. 1st, 2008|10:55 pm]
Lab Rats


So, first off, 'ello all!

Second, if anyone works with mice, perhaps you can help me with this question.

Do you know of any company (not R&D or PBL) that makes a good Interferon-alpha ELISA for mouse serum that is really sensitive? I just ran the R&D ELISA, know I did everything perfectly, and it stunk, royally. Thus I am frustrated at the lack of good ELISA readings (the standard curve wasn't even graphable!) and wondering if there is another good ELISA out there or if I need to start looking for other readouts for Interferon-alpha.

[User Picture]From: lickis
2008-11-03 02:10 am (UTC)
Sometimes ELISA is like that. The standards never actually make a straight line, it really is a curve. When you're at the low end the values can sometimes appear to be all over the place. If you're using a kit you tend to assume that they wouldn't tell you to do standards that are below the linear part of the curve because they'd be useless, but the thing is that especially when it depends on room temp for reactions the colour development isn't the same every time. So perhaps you needed to leave the development longer or something.
However, if that is the case it's basically irrelevant if your samples are at the same point in the curve. To get good results you would then need to dilute your samples less so they end up nearer the top half of the curve where it works nicely. That can be a total pain in the butt with mouse serum because you don't have of it (we need special mutant mice that have huge volumes of blood!). It can sometimes be helpful to half volumes and do a 50uL ELISA (assuming you usually use 100ul) so that you don't need as much of it. This has always worked well for me :)
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[User Picture]From: lickis
2008-11-03 02:14 am (UTC)
...you don't have a lot of it, that is. I should start proof-reading _before_ posting ;)
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