Methods and Techniques Discussion => Western Blotting (WB) => Topic started by: ghostdog on April 05, 2006, 09:19:49 AM

Title: Antibodies for western blotting experiments?
Post by: ghostdog on April 05, 2006, 09:19:49 AM
I have always wondered if a supplier states that an antibody is suitable for western blotting but does not state that it is suitable for another method i.e. immunhistochemistry does this mean it wont work ? if so why not? surely its the same epitope  ?

The reason i ask this is because i am trying to find an antibody and it states in nearly every supplier that it is suitable for western blotting but that it has not been tested on IHC.


any help or answers would be great

many thanks
Title: Antibodies for western blotting experiments?
Post by: ImmunoNYC on April 05, 2006, 09:46:04 AM
Antibodies that work in western very often do not work in IHC or flow as westerns often require that the proteins be denatured (although not necessarily always true). Since in IHC and especially flow we are looking at native (folded) proteins this can often cause problems. An antibody might be generated by a supplier because it specifically recognizes an antigen that is exposed when a protein is denatured. This antigen may be difficult to detect when the protein is folded in it's native state much less then fixed in a cross-linking fixative. (and vice versa for IHC specific Abs).

I find that when companies say it works in western without mentioning that it does not work in IHC, it usually means they have not tried it. IF you are desperate you can try it but it may or may not work.
Title: Antibodies for western blotting experiments?
Post by: ghostdog on April 05, 2006, 10:19:06 AM
that is brilliant thanks so much.
Title: Re: Antibodies for western blotting experiments?
Post by: FDossani on November 02, 2011, 05:32:22 AM
very well replied..i really appreciate that this matter is summed up in a good manner.
Title: Re: Antibodies for western blotting experiments?
Post by: AnneHowell on May 13, 2015, 02:37:53 AM
Yes , before you purchase you need to ask it clear about its applications. Most suppliers do note this at the productdescription on their website. You click the product name to enter the detailed description page and will see such information. By the way, if you want antibodies for western blotting, you can go there : http://www.creative-diagnostics.com/Antibodies.htm
Title: Re: Antibodies for western blotting experiments?
Post by: MMaggie on January 16, 2017, 08:06:12 AM
True, It may or may not work in different experiments.  :) It is always important to ask the distributor or supplier specific questions related to the work. Mostly, Kits and Antibodies are supplied in specific and sometimes custom made for specific use.