[Histonet] Differences between WB antibody and IHC antibody

[koellingr <@t> comcast.net]

TF and Emily,

You can use antibodies designed for Western blots.  They can work.  In a western blot, a lot of times these are run under denaturing conditions (a type of soap is used to break the secondary and tertiary structure) and so the epitope seen is linear.  That antibody for Western's works at a linear epitope (which is only a few small amino acids out of the whole proten structure).  Your tissue IHC is based on proteins that have not been linearized and are more native.  So if it happens that the antibody recognizes an epitope that happens to be linear within the non-linear protein, it certainly will work.  If it happens to be an epitope that is hidden away within the 3-D native structure and has not been linearized, it won't work.  There are WB that use (native) non-denaturing protocols so if the antibody works in that situation it might very well work in IHC.  There are protocols for placing tissue sections in SDS to try to emulate the conditions of a denaturing WB to be able to utiliz
e those antibodies, but these are not very standardized or widely accepted procedures.  Have used successfully WB antibodies on tissue IHC with almost no trouble.  Other WB antibodies I couldn't get to work on IHC despite heroic efforts.  Is all empirical; you just have to try.

Raymond Koelling
Research Scientist
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 
From: "Emily Sours" <talulahgosh <@t> gmail.com> 

> If anyone replies with posting to the list, please forward it. I've always 
> wondered the same thing. 
> 
> On Fri, Jan 2, 2009 at 8:15 AM, TF wrote: 
> 
> > Hi 
> > Just wondering whether the antibody for WB (only WB, IP on datasheet) can 
> > be used for IHC use? 
> > If not, why? 
> > 
> > Can I just increase the concentration for IHC application? 
> > 
> > I want to learn more about the underlying production processes. 
> > Thanks! 
> > 
> > 
> > 2009-01-02 
> > 
> > 
> > 
> > TF 
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> 
> 
> 
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