What would you recommend? It should be cheap and of constant / defined composition, comparable to defatted milk powder (NFDM).
Thanks!
If you really want to avoid animal origin, I have no idea. If replacing milk with something less complex is ok for you, casein is the blocking agent in milk and could be purchased in purified form. The only trick is to be patient for diluting it with gentle stirring, slowly adding the protein.
But again, for the original question (non-animal origin I ave no answer).
"SuperBlock" blocking buffer, a purified glycoprotein could be used for blocking. However, the source of this blocking glycoprotein is not given.!
I would recommend you to read this.
https://www.thermofisher.com/in/en/home/life-science/protein-biology/protein-biology-learning-center/protein-biology-resource-library/pierce-protein-methods/blocking-buffers-western-blot-elisa.html
--best regards
Odyssey Blocking buffer (non mammalian) is very useful
https://www.licor.com/bio/products/reagents/blocking_buffers/odyssey_blocker_pbs.html
Hi Wolfgang,
based on price and general availability you should investigate soybean peptone, soybean protein powder or similar products. There are also bulk purified single proteins such as alpha-amylase from barley available at somewhat higher costs. I guess its a try-and-error approach. Good luck hunting!
M
Markus, I thought about soy protein, too, but was concerned about lectins possibly causing undesired background.
Yes, soybean lectin may be a potential problem, but not necessarly a no go. First, if you use soybean peptone the lectin may be sufficiently degraded. Second, soybean agglutinin has a metal-ion binding site and thus may simply be inhibited by using EDTA. Third, lectin binding may be further suppressed using free sugars (Galactose, GalNAc). Fourth, the lectin probably is more specific to glyco-lipids and O-glycans and N-glycoproteins such as antibodies may not be recognized efficiently. So in the end i guess its worth a try.
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