I am interested to better understand the technical and practical implications of using a validated antibody for a specific immunofluorescence technique in another one that the same antibody is not validated yet.
I would say that the antibody approved for one immunofluorescent application would probably be able to be translated to another assay type. Are you trying to take to FACS from immuno(cyto/histo)chemistry? The main thing you can do is run a very small experiment for whatever new application you want to perform, while titering the antibody. I don't know what antibody you are looking at or how well it works. The company's recommended concentration is always lower than necessary, so start of there but include a dilution range in order to determine which concentration is best for the new application.
ABs validated for immuno can be used in flow cytometry, you will need to dilute appropriately, the other way around, i don't think so, it could work for western blot, but protocols may require modifications.
but you will have to try and see, if the company does not recommend an application, it means that it did not pass their validation standards for said application, if the data they acquire in their testing is ambiguous the application will not be recommended.
Hi Peres,
I'd say if the antibody is validated for Immunofluorescence,it has a high specificity in recognizing particular epitopes, and therefore it will be suitable for most of the other detection techniques. But this won't always happen other way around (esp when an antibody is validated for western blot transfer,most probably it has to be validated for IF seperately)
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Antibodies