Another Tech was interested in this subject so I'll forward this to the
entire list.
 
Glen
-----Original Message-----
From: Dawson, Glen 
Sent: Monday, October 14, 2002 2:50 PM
To: 'Van Eyck, Deb'; 'http://ls.cytopathnet.org'
Subject: RE: 


Deb,
 
I made a Melanoma Cocktail for the Medical College of Wisconsin using
Mart-1, Melan A & Tyrosinase from 3 different vendors.  I first optimized
each antibody individually and then made the cocktail with the best
individual dilutions in mind.  Keep in mind that you cannot simply add the
working dilutions of each separate antibody together to get the cocktail as
this would be too much diluent.  Instead you must add concentrated antibody
to a designated amount of diluent.  Example:  Optimal dilutions are:  Mart-1
(1:500)  Melan-A (1:100)  Tyrosinase (1:50).   To 9.68 ml. of diluent, add
100ul of Melan A, 200 ul of Tyrosinase, 20 ul of Mart-1 to obtain 10 ml of
the working Melanoma Cocktail.  
 
As for species, as long as the secondary antibody accounts for all the
species the concentrates are made in, it should be fine.
 
Charging for this antibody doesn't account for the added expense of having 3
antibodies in one but it works so well for the patient that my lab just
takes it in the shorts on this one.  I don't know what kind of reporting
issues the cocktail causes, maybe a nice pathologist out there can help you
out on that one.
 
Glen Dawson
Milwaukee, WI

-----Original Message-----
From: Van Eyck, Deb [mailto:deb.vaneyck@phci.org]
Sent: Monday, October 14, 2002 12:14 PM
To: 'http://ls.cytopathnet.org'
Subject: 



Here is another question for all you immuno wizards.  At several of the
immuno workshops at the NSH I heard several people say they made up antibody
cocktails by mixing equal amounts of optimally diluted antibodies.  (For
example HMB45, Tyrosinase and Mel A or Mart 1)---Not too worry proteins seem
to like each other!  In the immuno world this just seems a little too easy,
plus you can charge for the use of the separate clones?  How would you
report this????? Please explain ---aren"t there some rules like same
species, same diluents, etc?