alfandar at vasci.umass.edu
Mon Jun 27 13:38:45 EDT 2016
My R24 to produce at least 100 monoclonal antibodies to Xenopus protein obtained a very good score likely to be funded sometime in october. This proposal is based on 2 approaches.
1 Immunization with Xenopus embryo extracts (Stage 22 membranes and multiple sub fractionation) to produce antibodies that will work on endogenous proteins but without specific protein target. These will be ultimately identified by mass spec. We have produced our first one already against Ribophorin 1 (rpn1) that turns out to be a fantastic loading control and an excellent marker of the TGN by IF.
I fully expect to get some antibodies to Yolk (Vitellogenin) that could be useful to clear ms/ms samples using covalent beads. These will be available as soon as the characterization is done.
2 Immunization with specific proteins that the community request. In order to get that list (I already have a preliminary list). I will need to obtain
1 why is this protein important for the community.
2 a convincing evidence that antibodies against other species do not work (Low conservation)
3 a Flag tagged construct in CS2 or other plasmid with a CMV promotor.
It is much easier to make antibody against cytoplasmic or nuclear proteins so I will likely start with those. I also plan to make antibodies specific to growth factor receptors.
All of these will be distributed via the DSHB as soon as the characterization is complete.
Feel free to contact me about your favorite proteins. I can start before I know if I have the money but I am sure I will have the yolk mAb before that.
Dominique Alfandari Ph.D. Professor,
MCB graduate program director,
Dept. Of Veterinary and Animal Sciences
University of Massachusetts, ISB rm 427B.
661 North Pleasant Street, Amherst MA 01003
Ph: 413-577-4269 Fax:413-545-6326
When the Fathers let the children do anything. When the Sons do not care about their words,
When Teachers tremble in front of their students... When finally the young disregard the laws,
because they do not recognize any longer, above them, the authority from anything or anyone...
Then, in all its beauty and its youth, is the beginning of Tyranny.
Platon 428-347 BC.
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