Immobilization of Beta-glucosidase with glutaraldehyde

[Lindsayyyy]

Hello everyone,

I want to immobilize beta-glucosidase (on Sepabeads with an amine group). The problem I have is I don't have a strong chemical/biological background. The immobilization should happen with glutaraldehyde but I can't find an appropiate paper with a protocl I can use. In every paper I found there are always insufficient information for me in order to reproduce it.
Does anyone have a good protocol or a paper where I can find something about this? I'm quite desperate at the moment because I've been trying to immobilize this enzyme for quite some time now without any success.

Thanks for your help.

Lindsay

 

[Ygggdrasil]

Glutaraldehyde is a fairly strong crosslinking reagent and might inactivate your protein during the crosslinking reaction. A potentially better way to crosslink would be to use carboxylated beads, treat activate your beads with EDC and NHS, then react your protein to the beads. The following site has some good resources for protein crosslinking and some kits biochemists commonly use for protein crosslinking: https://www.lifetechnologies.com/us...thods/carbodiimide-crosslinker-chemistry.html