I am trying to hydrate CNBr activated Sepharose 4B using 1mM HCl, the resin was expired 10 years ago and it was purchased from Amersham Bioscience, Sweden, I am trying to get catalogue and details through their website but unfortunately, it's not available.
The problem is it's not getting hydrated, ideally, the 0.25gm of resin should swell up to 1ml but it's not. Should I use this resin? Any suggestions?
Hi Ahmad, I agree with Omar in that if the resin was wet at anytime in storage , it is probably degraded and useless
But CNBr is a potent antifungal and antibacterial, so it might still be usable.
Do a quick and easy experiment to see if it is still useful.
Take any old Ab-HRP conjugates you have in the fridge (that are not in tris or stabilized with BSA or other carrier proteins). Incubate it with a sample of the old CNBr Sepharose and wash it like you would do with fresh resin.
Add HRP substrate to the resin. If it turns blue, then your old resin is probably OK.
Thanks Omar and Stein, the resin is dry and was well stored in a fridge.
I will give it a try with HRP
Hi Ahmed
The resin is still available from GE lifesciences - in the UK at least. I have attached the product literature.
In my experience a 1:4 swelling ration is a bit high and it is usually 1:3-3.5 ml depending on how well it is freeze dried at the manufactuerer. If the resin wasn't in a heat sealed foil bag then it may have abosrbed some moisture already, if it has it will swell less and be less active.
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