Protein elution from amylose resin (View forum version)



spellberg56

Posted 18 March 2010 - 07:08 AM

Hello, I have my protein of interest bound to Maltose Binding Protein (MBP). I have bound this fusion to amylose resin in a 1.5 ml microfuge tube. I have already tested my amylose bound samples to ensure they are present. The protocol says to elute with 10mM maltose in buffer but the protocol also uses a amylose column, not a microfuge tube. If I add the maltose to the amylose bound resin, do I remove the supernatant to get my eluted protein after spinning down? I won't be able to take fractions when using a micorfuge tube.

Thank you

mdfenko

Posted 19 March 2010 - 07:36 AM

you are performing a batch elution. no problem, just make sure that you thoroughly wash the resin before you elute or your product will be contaminated with material that didn't bind.