I have a secondary and two primary antibodies in my lab (which can be used against actin and HSP70 in Western blotting). The application notes in their data sheets only says that they can be used for Western Blot but doesn't mention if they can identify the protein in denatured state (because of SDS). Can anyone tell me if antibodies in general can identify the SDS-denatured protein. Can I use them for my studies?
1. If the datasheet of an antibody says that the antibody works for Western blots (WB), then it means this antibody can recognize denatured protein.
2. If the datasheet of an antibody says that the antibody can be used for IP, Flowcytometry (FC), IF (not paraffin but frozen such as OCT) but not WB, then this antibody recognize only confirmation specific epitopes and will not detect denatured proteins in WB.
3. There are certain antibodies which works for both WB and IP/IF/FC depending on the single chain hydrophilic epitopes (on exterior surface or protein in 3D which is also exposed in denatured conditions).
4. Finally the most important thing..!! Certain companies (I am not mentioning company names here) list that an antibody works in WB/IF/FC/IHC-P/IHC-FFPE and whatever applications possible.. but might not work for most of these applications. Its nothing but marketing strategy to increase sales..!!
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