WANTED: help w/Drosophila embryos

ylevy at binah.cc.brandeis.edu ylevy at binah.cc.brandeis.edu
Thu Jan 19 11:48:32 EST 1995


Dear Net Folks,

I'm intersted in doing an immunofluorescence localization pilot experiment
with Drosophila syncytial blastoderm embryos.  Specifically, I am studying
a centrosomal antigen that appears to be ubiquitous in Eukaryotes and
would like to know if it is present in Drosophila centrosomes.  I get a
positive reaction to embryo proteins on Westerns with my antibody.

As we are not a fly lab, I initially obtained fixed 0-3 hr embryos from
another lab here at Brandeis University.  They were formaldehyde fixed,
dechorionated/devittelinated and in MeOH/EGTA (i.e. ready for rehydration
and staining).  I did several experiments varying antibody incubation
conditions (such as temperature, concentration, and time) but every
experiement yielded the same poor result: general fairly high level
uniform fluorecence.  The same result was obtained with monoclonal
antibodies against alpha- and beta-tubulin. 
 
Does anyone with experience staining early Drosophila embryos have any
ideas what can be wrong?  I was struck by the completely uniform signal. 
Not even a hint of differential antibody binding.

Also:  if there are any Saints out there willing to send me some fixed
embryos (preferably by the procedure of Warn & Warn (1986) I'd be happy to
pay the shipping charges and would be forever grateful.


Thanks much, sincerely,  Yaron.


Yaron Levy
Dept. of Biology
Brandeis University
Waltham, MA 02254

T 617-736-3152
F 617-736-3107

ylevy at binah.cc.brandeis.edu



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