Author Topic: IHC antibodies and WB  (Read 6458 times)

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Offline dustman

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IHC antibodies and WB
« on: February 15, 2008, 06:04:58 AM »
There are several anti-COX2 antibodies on abcam list that are sited to work in different IHC techniques but not in western blotting. Is it really possible? I thought almost all epitopes are exposed after treatment with mercaptan and SDS.

IHC antibodies and WB
« on: February 15, 2008, 06:04:58 AM »

Offline CanuckPhD

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Re: IHC antibodies and WB
« Reply #1 on: February 15, 2008, 10:29:24 AM »
Sometimes antibodies that work on WB don't work with IHC. However, if the antibody is recognizing a non-linear epitope on the protein it may not work on WB but give staining with IHC. This is because proteins are often boiled, put in SDS etc for WB, which will remove the tertiary structure. This may then destroy the non-linear epitope that the antibody is recognizing.

Offline dustman

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Re: IHC antibodies and WB
« Reply #2 on: February 18, 2008, 02:58:10 AM »
Thank you.

Although I'm bit unsure if this applies to this case. Immunogen is stated to be synthetic peptide (C terminal, rat) and I'm not sure if relatively short peptide can have complex tertiary structure. Again, immunogen is *considered to be commercially sensitive* and as such it's hard to say that for sure.

Re: IHC antibodies and WB
« Reply #2 on: February 18, 2008, 02:58:10 AM »