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How to select a primary antibody?

When looking for a primary antibody, one will first type his target name in any search engine. But then how to select among the different available commercial antibodies?

Question: How to choose between a monoclonal and a polyclonal?

Answer :

 

Question : Which host will show the best results ?

Answer : Unfortunately, there is no rule that some animal species will always give better antibodies. This is dependant on each antigen and even in some cases on each animal: the quality of the antibody depends on an immune reaction which is also oriented by the immunized animal history and genetic back-ground.

However, the host choice is critical for multiplex labellings where you need to discriminate each antigen by a differently labelled secondary antibody. The primary antibody should be selected from a species as different as possible from the sample: this will avoid part cross-reactivity of the Secondary with the sample. This secondary must be cross-adsorbed against the other primary species, and against the sample species if this one can show endogenous immunoglobulins (eg liver, thymus, skin...).


Question: Why should I consider a Rabbit monoclonal antibody?

Answer: Rabbit hybridoma partners are now available for mAbs development. They allow the release of Rabbit mAbs which usually show diverse epitope recognition, high specificity and affinity, exceptional immune response to smaller epitopes & greatly improved antigen recognition.

Question: How to select an antibody if my application is not guaranteed?

Answer : you can select the most likely to work through the antigen configuration needed: WB and IP work on denaturated proteins. ELISA, FCM, IF and IHC (without fixation) are compatible with native configuration proteins. Biological activity is most likely to be found in pAbs raised against full-length proteins…