Question for the lab geeks out there regarding general methods for antibody detection of specific analytes in cells, serum, and urine.
Do you have a favorite book, chapter, or any other reading material that you would use to guide students through designing and validating their own ELISA for a serum or urine protein? If you have any lecture slides you use and could share, I’d be happy to credit you. Some famous guy(s) gave me his H1N1 slides last year and helped me look like a genius.
I know that it takes a mighty, mighty fine antibody to do this relative to one for an immunoblot or even an in-cell western.
Of course, I know that one of my advisors would tell me to just buy a damn kit but that is the wrong answer for my current needs.
And no, I don’t have the money for a Luminex xMAP thingie just yet.
For those of you who wonder what the heck I’m talking about, ELISA stands for enzyme-linked immunosorbent assay. It’s a method that uses a highly-specific antibody to measure stuff in any biological matrix – applications include HIV testing, PSA levels, and even home pregnancy tests. Most of the time ELISAs are used to measure proteins but they can also sometimes be used to measure small drug molecules such as the cardiac glycoside, digoxin (a drug taken by someone with congestive heart failure).
In fact, here is a link to a nice animation I found on how a home pregnancy test works.
The Virtual Immunology Lab at the Howard Hughes Medical Institute (HHMI) site is neat but I’m wanting to present more technical detail on assay development and validation.
In the meantime, thank you for any advice.
Photo credit: New England Biolabs