ELISA protocol request - (Dec/26/2008 )
I already went through the protocols, and couldn't find what I was looking for. If anyone can help me, I'd sure appreciate it.
I'm a bit of a kludge at the molecular stuff; I mainly worked on transgenic plants, so bear with me. What I'd like to do is look for antibodies in blood sera to a specific bacterium. I can grow the organism in pure culture, no problem. But I could sure use a top-to-bottom protocol for ELISA detection of antibodies in sera to a given bacterium.
Anyone have one handy? Thanks in advance!
Reagents you will need are:
1. Purified ag from the culture (or can you obtain commercially?)
2. Negative and positive sera samples (confirmed by a reference method)
3. Enzyme labelled polyclonal or monoclonal ab against human IgG (post infection detection) or IgM (recent infection). (also commercially available)
Take a look at some of the commercially avaialble kits for detection of abs to bacteria (such as salmonella, etc). This will give you some understanding as to the test method you will be developing.