RE.Negative control on IHC's

From:"Brennan, Liam" <>

My reasoning on this issue is fairly simplistic, if you put up a positive
control slide then not all elements in that control tissue will stain
positive with the antibody being used therefore the non-staining areas are
your negative control. This is further backed up by the non-staining areas
of your Test section acting as a negative control.
	From a pure science point of view, you could run a multitude of both
positive and negative controls, with regard to reagents, antibodies, tissues
etc. and you might possibly end up with a ratio of 6 or 7 control slides per
test slide. This although scientifically correct(covering all possible
outcomes), is just not practical in the setting of a diagnostic lab. where
the workload just would not permit it, as well as the financial implications
on the cost per test.

Liam Brennan
Belfast City Hospital

> >Is there a good reason to run a negative control on every block when
> >staining IHC's?  We do a lot of keratin stains on lymph nodes from the
> >same specimen (not part A, part B, etc.) and we run a negative control
> >along with the keratin on every block.
> >
> >Laurie Colbert
> ********
> Speaking at least from research point of view:
> Officially, every tissue block needs a negative control using a non-immune
> reagent (normal rabbit serum, fake monoclonal antibody) replacing your
> primary antibody. This non-immune control should be applied in the same
> immunoglobulin concentration as the primary antibody.
> See for example JHC 48:1431-1437, 2000 Table 1. Most immunohistochemical
> journals do request for such non-immune negative controls.
> Chris van der Loos
> Amsterdam, The Netherlands

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