Chromatography Forum

Hi,

I have lyophilized protein and I would like to solubilize them. I want to test them on a silica based column with a graft protein inside.
Do you know how to solubize them?

Thanks.

You will need to know a couple of other things to determine the correct way to get them into solution. First, what where they lyophilized in (solvent-wise)? If they were lyophilized in a buffer that they are soluble in, then those buffer salts are still present and all you need to do is add water. Warming, stirring, and possibly filtration may be necessary depending on how well they were lyophilized. If no buffer salts are present, you will need to dissolve them in a suitable buffer. In some cases substances (sugars for example) are added before lyophilization to preserve the structure of the protein by taking the place of water molecules. You may need to add a dialysis step to remove them.