Immunoaffinity Subtraction

Amy Walker agenbite at asterix.bio.sunysb.edu
Mon May 8 23:01:42 EST 1995


In article <christer.ericsson-0805952010360001 at drop-dead.cmb.ki.se>,
christer.ericsson at mog.ki.se (Christer Ericsson) wrote:

> In article <3ohqli$795 at worak.kaist.ac.kr>, dykim at bioneer.kaist.ac.kr
> (Dooyeon Kim) wrote:
> 
> > 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
> 

Hi, I have read some stuff about using "immunosubtraction'' to develop
antibodies specific to certian stages of hematopoiesis.  The idea was to
make polyclonal sera against one cell type, and absord the sera against
the cells you want to exclude,  before going on to make monoclonal
antibodies.  I realize you are after something a little bit different, but
maybe there's information  you can use in these papers. Anyway the best
references I have are

1) Kornfeld S. (1983) Cell Surface antigens on avian erythroid cells:
their expression at various stages of differentiation. Expl Cell Res. 150,
p152

2) Kornfeld S., Beug H., and Graf T. (1983) Detection of avian
haematopoietic cell surface antigens with monoclonal antibodies to myloid
cells: their distribution on normal and luekaemic cells of various
linneages. Expl Cell Res. 143, p383

Good Luck,
Amy Walker



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