Titrating the antibodies

Michie, John, Dr <jm5@sun.ac.za> jm5 at sun.ac.za
Fri Jan 18 03:17:49 EST 2008


There is a tutorial (and sample files) by FlowJo available at

 http://www.flowjo.com/home/tutorial.html

 

It takes you through the steps of titrating a FITC labelled anti-CD8
reagent to determine the most economical concentration for staining.

 

Kind regards

John Michie

   +27 21 9389 539

 +27 21 933 8886

   083 449 2225

 

-----Original Message-----
From: Abdul Khan [mailto:khan.abdulh at googlemail.com] 
Sent: 17 January 2008 12:10 AM
To: cyto-inbox
Subject: Titrating the antibodies

 

Dear All,

 

Is it necessary to titrate all the antibodies before doing the FACS
analysis, eventhough most of my antibodies are pre-titrated/pre-diluted
(as per datasheet) but some of them say only concentration like 0.2 mg.
I have got 15 antibodies and would appreciate any help/advice in this
regard. Can anyone please suggest the best way to titer the antibodies,
I will be doing 3 color flow cytometry. 

 

I would also like to know if it is best to buy the antibody and isotype
control from the same company ? I bought one antibody from a different
company but I already have the isotype control from another company, now
is it recommended and worth buying the isotype control from the same
company from where I bought the antibody or shall I buy one ? 

 

I shall look forward to your responses. Thanks a lot.

 

 

Best wishes,

 

Abdul

 

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