What is rocket electrophoresis?

David Johnstondaj daj at uk.ac.ic.nhm
Fri Aug 27 03:56:17 EST 1993


On 26 Aug 1993 16:05:47 GMT,
  Vivian Miao writes:
>
>OK, I'll ask ....
>
>What's rocket electrophoresis?
>
>
Its a technique for quantifying proteins (or anything else you can raise an 
antibody against). If you are familiar with SRID (single radial 
immunodiffusion) assays, its basically similar in concept but involves 
electrophoresis.

Basically you make a thin, horizontal agarose gel containing your 
antibody and cut a row of small circular wells across the middle of it. You
then place your samples and series of standards of known concentration in 
the wells and electrophorese them (in a LKB multiphor or similar tank 
using paper wicks dipping into buffer solutions). As the antigen migrates 
through the gel it contacts antibody and at a certain ratio you get the 
classic antibody-antigen precipitin reaction occuring. The more antigen you 
have, the further it migrates before the precipitin reaction goes to 
completion . You end up with an elongated elipsoid precipitin reaction with 
a rounded end at the well end and a sharp point at the limit of migration. 
This is basically "rocket shaped", hence the name. The gel is 
usually then dried and the precipitin line stained with coomassie 
blue. The length of the rocket is proportional to the concentration of 
antigen which is determined by reference to a plot from the standards.

A useful and fun technique from the archives of science. It used to be used 
a lot in clinical chemistry before the advent of modern analysers.

Cheers

DAJ
David A. Johnston
Dept of Zoology, The Natural History Museum, Cromwell Road,
South Kensington, London SW7 5DB.
(tel 071 9389297, fax 071 9388754, email daj at nhm.ic.ac.uk)



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