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Next Subject		 dialyzationqianxm2001-09-19Click here to register.
Dear Sir,
I have prepared the loposome ,which is 200-400nm using the Avanti extruder. In the system, I want to make the Con A protein to be within the liposome. Now I found that the Con A protein exists in two ways. First is free in the solution, and the other is encapsulated in liposome.
The separation of the free Con A and liposome is my work in next step. The methods are centrifugation and dialyzation, I think. Would you please give me some advise, or tell me which membrane or centrifugation filter I can book in your company to separate the free Con A and liposome(200-400nm)? I seek the information in your catalog and web-side all the afternoon, but I could not have the last decision.
The molecular weight of concanavalin A is 27,000.This size molecule in true solution should pass through a 0.2 micron pore.

Xinming Qian
My mail address: S71, Apt6, University place, Stillwater, OK 74075
(Department of Chemistry,
Oklahoma State University,
Stillwater, OK 74078)
Tel: 405-3320815(H); 405-744-7307(O)
 
Re: dialyzation
dave2001-09-21Click here to register.
200 nm corresponds to an MWCO of about 2,000,000. Your separation (27,000 from 2,000,000) is a relatively easy one so you have lots of choices that depend upon your sample size and how often you plan on doing the experiment.

Dialysis with a 300,000 or so MWCO membrane will work.

I don't know how strong your liposomes are. If they can take the abuse, you can certainly use a similar MWCO centrifuge filter (small samples) or an regular ultrafiltration system (larger samples).

You probably want to be sure that the Con A cannot diffuse through you liposome's membranes (I have no idea if it can or not). If it can diffuse through you will always have some free protein.

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