di58 at pop.th-darmstadt.de
Wed May 10 06:58:02 EST 1995
In article <3ohi8v$1q5 at worak.kaist.ac.kr>, dykim at bioneer.kaist.ac.kr (Dooyeon Kim) says:
>I have some ideas on the detection and purification of newly expressed protein.
>Instead of using cDNA subtraction or 2D PAGE analysis, I will use immunosubt
>ractive methods. The princeple of this method is producing the polyclonal
>antibodies against whole protein mixture of tissue or cultured cell and select
>ing the antibodies against newly expressed protein by passing the antiserum
>against the protein(antigen) column of control tissue of cell line. After immuno
>subtraction, the newly expressed protein will be easily purified by immuno
>precipitation and detected on SDS PAGE gels. Dose anybody advice me about this
>idea? Any Comments will be greatly appreciated. Thanks for reading
>KAIST, Deajon Korea. Dooyeon Kim ......................
> dykim at bioneer.kaist.ac.kr
instead of the proposed laborious "cascade" technique try an immunotolerance
protocol first published in J.Immunol.Meth. Vol.100, pp73-82 (1987). We used
it two times with reasonable success.
More information about the Methods