Dear colleagues,
I have a question about epoxy activation - is it possible to use 60% technical grade 1,4-Butanediol diglycidyl ether instead of 95% to immobilize proteins on a Sepharose beads simply by adding proportionally more? Will there be problems due to the purity of 1,4-Butanediol diglycidyl ether? It’s just that the cost of technical grade is much lower - at Sigma Aldrich 250 g of 95% purity 1,4-BDGE costs $693 and 500 ml costs $169.
Is there any specification given regarding the remaining 40 % of the technical product? Typically, the proteins to be immobilized a rather expensive and especially if the technical product is uncharacterized, I would not use it for the suggested application. Is there another vendor who may offer smaller amounts of the compound or do you really require 250 g or 500 ml?
Dear Martin and Lorin,
Thank you for so quick reply! I also thought that contaminants can affect the performance of the affinity resin. But I needed to hear confirmation, thanks for the valuable advice!
I thought to make an affinity resin with a spacer, in principle, the option of activating with epichlorohydrin and immobilizing 6-aminocaproic acid acid, as a spacer, and coupling on the carboxyl group with carbodimide also works. But I thought to find an easier way.
Dear Rafil,
do you require the additional spacer and what protein should be immobilized on the resin? I had good results with Cys-tagged Proteins on epoxy coatings but I believe NHS-activated carboxyl groups or aldehydes ( from acidly opened and oxidized epoxides) may immobilize proteins faster.
Dear Martin,
I want to immobilize a ZZ domen of protein A on a resin, I have not tried it with a protein yet. But everything worked with one enzyme inhibitor.
For the activated resin performance checking I use that approach: small amount of the activated resin incubate with rhodamine C. The dye is bright and immediately visible it is immobilized after washing.
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