RE: in case of isotype control - substitute?

From: Candace Enockson (enockson@musc.edu)
Date: Tue Oct 31 2000 - 13:03:08 EST


I would agree with Greg.  If you have a real negative control you don't
need an isotype.  Often isotypes are not true negatives, that is, they
don't mimic the non-specific binding of the antibody, even tho' they are
isotypically matched.  When you have a negative population you can use,
that is better than an isotype.

Candace Enockson
Medical University of South Carolina

--On Tue, Oct 31, 2000 7:49 AM +1030 "Hodge, Greg (HAEM)"
<hodgeg@mail.wch.sa.gov.au> wrote:

>
>
>
>> ----------
>> From:		Maciej Simm
>> Sent:		Tuesday, 31 October 2000 1:17
>> To:	Cytometry Mailing List
>> Subject:	in case of isotype control - substitute?
>>
>> Maciej,
>>
>>	I dont bother with isotype controls for IL-10 production by
>> monocytes, I find unstimulated monocytes do not make any IL-10 and so use
>> this as my "negative" control.
>>
>> Greg Hodge PhD
>> hodgeg@mail.wch.sa.gov.au
>> ------------------------------------------------------------------------
>> -- ----------------------------------------------
>>
>> On 31 October, Maciej wrote-
>>
>> Howdy Everyone,
>>
>> I am running IL-10 in human monocytes using a PE conjugated Rat anti
>> Human IgG1 antibody. There is a shortage of intracellular isotype
>> control for that. What can I use instead? someone recommended using
>> anti rat IgG2 etc.. but I've read on here earlier the 2's are picked
>> up by monocytes..
>>
>> so how about a mouse IgG1?
>>
>> Maciej
>>
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