Estrogen Receptor Assay

Raymond Pierre raymondp at ERE.UMontreal.CA
Wed Oct 30 00:04:06 EST 1996


Does anyone have experience with the assay of estrogen receptor?  We are 
trying to measure the binding capacity of estrogen 
receptor found in fish liver. We would also like to do competitive 
binding  study with several potential xenoestrogens.
	The assay method  used H3-estradiol as the ligand, 
diethylstylbestrol (100 X in excess) to discriminate for specific binding 
and Dextran-Activated charcoal preparation to separate estradiol-receptor 
complexes from free estradiol in the incubation mixture.  Those familiar 
with the Scatchart plot determination of estrogen receptor will 
recognize the method.
	We have been trying to measure the amount of estrogen receptor by 
this technique which looks fairly simple  but  without 
success for several week now 
	As a positive control (fish from which estrogen receptor should be at 
their maximum) we used female trout at a stage representing about one month 
before spawning. So we are pretty sure the receptor are there.  We tried 
several modifications to the technique of isolation and that of binding 
assay (pH of buffer, concentration of glycerol etc)
	If anyone out there knows of limitations and/or precautions regarding 
this technique, we will greatly appreciate hearing from you.

Thanks in advance,

Pierre




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