Re: Antibody Cocktailing

Hi Sara,

Why are you not getting BD to assist you?

Regards

Martin J Dominguez
Cellular Solutions Specialist
Beckman Coulter


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	     "Markestad, Sara"						   
	     <SMarkestad@sjha.						   
	     org>							To 
				       "Cytometry Mailing List" 	   
	     20/07/2005 18:58	       <cytometry@flowcyt.cyto.purdue.edu> 
									cc 
								   Subject 
				       Antibody Cocktailing		   




Hello Flowers!


	We have recently purchased a BD analyzer and are attempting to go
to 6-Color Leukemia/Lymphoma Panels. The problem we are running into is
that the 6-color antibody cocktails that are out there are not compatible
with our panels and pathologists. We want to purchase individual antibodies
and create our own 6-color cocktails. Since this will be my first time ever
attempting anything along this line, I am hoping that there is someone out
there who will be able to lend some assistance.


Here is what I am thinking I will have to do:


	I am going to have to purchase the antibody in the colors and the
antibodies that I want. I will then have to run the antibodies individually
and get a mean channel. I will then need to add 1:1 concentration of each
antibody and then run that “cocktail” and see where the mean sits for each
antibody in the “cocktail”. I will then need to adjust the concentration of
each antibody till I get the correct mean channel. Once I figure out the
concentration of each antibody I will have to run that “cocktail” using
several amounts. (For example: 10ul, 20ul, 50ul, and 100ul) With these
amounts I am going to have to compare the positive population versus the
negative population and make sure I am using the correct amount.


This appears to be a lot of work but am I on the right track? If not can
someone please help me?


Any comments or suggestions would be greatly appreciated =)


Thank You in advance.





Sara Markestad, MT (ASCP)


St. Joseph's Hospital of Atlanta


404-851-7744


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