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Which isotype control should i choose


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#1 leyshmanioz

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Posted 14 January 2013 - 11:04 PM

Dear all please help a FACS noob,
I need to choose isotype control for my surface marker staining. Should i choose isotype control depending on the species i'm labeling, so if i have Ab against human CD44-PE conjugated my control should also be anti human IgG. Also does the choice of type of Ab depend on the specific stained one. For example if my Cd44-PE was IgGk then isotype control should also be IgGk and what is the difference between normal IgG, does it really make a big difference.

Thanks

#2 Tabaluga

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Posted 15 January 2013 - 04:30 AM

No, you choose the isotype control according to the species the antibody was raised in. For example, is your anti-human CD44 antibody made in rabbit ? If so, you need rabbit IgG (and your isotype control is not "anti" anything). Since your antibody is primarily conjugated to PE; you need a conjugated rabbit IgG-PE as isotype control. Ideally it should also be the same subtype (IgGk, in your example).

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