determining titer

[Ed Rybicki]

Caroline Szymeczek-Seay wrote:
> I'm producing antibodies in rabbit from synthetic peptides.  It's
> rapidly becoming time to check the Ab titer from the first bleed, and > realize that I have no protocol for this.  Any suggestions on protocol
> books?  I'm more of a DNA/RNA person, and haven't ventured into Ab
> production before.  Help will be appreciated.

See if your peptides stick to an immunoassay plate - then do simple
indirect ELISA (eg: stick a concentration range to the plastic [ and
KEEP the stuff if peptide is precious], and test a range of Ab
concentrations with it, and a non-reactive Ab as well).  Simplest things
often work best...!  Or do an inhibition assay, wherein you bind Ab to
peptide in suspension, then test reaction of Ab with native Ag.

-- 
                     Ed Rybicki, PhD  
      Dept Microbiology     |   ed at molbiol.uct.ac.za   
   University of Cape Town  | rybicki at uctvms.uct.ac.za
   Private Bag, Rondebosch  |  phone: x27-21-650-3265
      7700, South Africa    |   fax: x27-21-689 7573
    WWW URL: http://www.uct.ac.za/microbiology/ed.html      
                                        
    "Out here on the perimeter, there are no stars..."