We're thinking about using a monoclonal antibody (IgG) in an
attempt to immunolocalize a water-soluble molecule (MW <200) that
presumably exists in the cytosol or vacuole of plant cells. We think we
can get the McAb into the cells OK, then cross-link the McAb-hapten
complex using a fixative to another protein (or other structure), and
follow with a secondary anti-IgG `reporter' antibody. We know that
we cannot fix the hapten prior to introducing the McAb because it would
make the epitope unrecognizable. Washing away unbound secondary antibody
should not be a problem; however, I can't think how - prior to the
cross-linking step - we can wash away unbound primary McAb without
concomitantly losing the McAb-hapten complex. If anybody has experience
with this I'd appreciate receiving advice.
**********************************************************************
Rod Savidge, PhD, Professor | E-mail: savidge at unb.ca
Faculty of Forestry and \|/
Environmental Management \ | / Phone: (506) 453-4919
University of New Brunswick _\/ | \/_
Fredericton, NB CANADA \|/ Fax: (506) 453-3538
E3B 6C2 |
