Immunoaffinity Columns

[RoundUp]

Emma,
The ratio between ligand and gel is rather crusial since you might get 
over coupling if you use to little of the ligand. This means that a lot 
of ag binding sites might get bound to the matrix. When we do couplings 
we try to keep the ratio as close to 1:2 as possible, and dont forgett 
the blocking of unbound sites after the coupling. After coupling we check 
the coupling solution for protein content and reactivity with secondary 
antibody on a dot blot, it might be a good idea to do this with a sample 
of the gel after blocking unbound sites also to check the binding.
Lars.S at Plantbio.lu.se, Plant biochemistry, Lund University
_________________________________________________________________
 
"I was afraid of a united Church;it makes a mighty power,
 the mightiest conceivable, and then when it by-and-by
 gets into selfish hands, as it is always bound to do,  
 it means death to human liberty,and paralysis to human thought" 
                 Mark Twain