Immunoscreening cDNA expression library

[Geoffrey Kidd]

Back in the Stone Age, I did such a task.  I also walked to school 10 
miles in the driving snow barefoot and hungry, but that's another story. 
 I used a biotin/streptavidin ket called Vectastain, by Vector Labs I 
think.  I haven't heard of them since then (the mid-80's), but they 
could still be around.  I had no problem with it - worked like a charm 
the very first time, astoundingly.  I pre-adsorbed out the non-specific 
anti-E. coli antibodies first.  I don't recall now if the assay was 
peroxidase- or phosphatase-based, but since peroxidase self-bleaches 
after a short time, I'd go with phosphatase.  Either way, I'd avoid 
125-I because I find pleasure in avoiding the red tape of ordering, 
storage, cleanup, and disposal of radioactive materials.  Hope this 
helps.

Geoffrey Kidd

Snyder Research Foundation Staff wrote:
> 
> Dear Group,
>         I am thinking about screening a cDNA library with an antibody we
> have for a protein which cross reacts with the antibody.  I would like to
> hear from people who have actually done such screening so that I know some
> of the pitfalls I may face before they hit me in the face.  Also, does
> anybody have any recommendations about commercially available kits, and
> thoughts about 125I versus Alk. Phos'tase (the two kits I have found are
> both alk. phos'tase based).
> 
> Thanks,
> Ron Tate
> Snyder Research Foundation
> 
> (if responding by e-mail please put my name in the subject field)