Scientist Solutions: Life Science Discussions
    
Home » Forums » Antibody Based Technologies » Assay Development & Protocols » How do you microdialyze your protein?

Thanks to our sponsors who make this site possible

How do you microdialyze your protein?

RSS Feed

Would you like to save this topic, event, protocol or job so you can find it again easily?

Just click the "Save to My Lab Drawer" link and the item will be saved in the My Lab Drawer section of your bench space.

Available to members only. Please log in or register for your free account now.

Chemgod1
United States

Send PM
See Mini bio

Status: Frog Egg
Frog Egg
Topic Started by Chemgod1
on 8/7/2008 11:43 AM   
Reply to this post Go to the top of the page

Hi I have this antibody that I need to run on gel. But it's been purified under high salt conditions (0.5M NaCl). I know that the gel will give me problems if I run it under the high salt. However, I have been told I can microdialyze my protein. but how??


Replies
samm
United States

Send PM
See Mini bio

Status: Cell & Tissue Culture, Antibody Technologies Moderator
Frog Laureate
Posted By samm
on 8/8/2008 14:05 PM   
Reply to this post Go to the top of the page

Pierce makes microdialysis cassettes.

As for your actual problem, samples with high salt tend to be more conductive. Thus, when sample salt concentrations are way higher than the gel salt concentration, current selectively flows in the region near the sample, rather than uniformly throughout the get. Since migration depends on the current flow, samples will run faster in regions of high current flow, in this case in the region containing high salt samples.

This can be simply solved by dialyzing your sample prior to loading OR by making a gel with a higher salt concentration. High-salt gels run warmer, and may need to be run at a lower voltage than a normal gel (active cold circulation, or running it in a cold room also helps).


Last edited Aug 08, 2008, 15:06 PM by samm

samm
United States

Send PM
See Mini bio

Status: Cell & Tissue Culture, Antibody Technologies Moderator
Frog Laureate
Posted By samm
on 8/15/2008 9:26 AM   
Reply to this post Go to the top of the page

If you don't want to get the Pierce microdialysis system, I just found a brochure for a Novagen D-Tube Dialyzer - it looks like a tube with windows where the dialysis membranes are. Also, we routinely use the 500ul Pierce dialysis cassette - that can simply be placed in a styrofoam holder, and kept in a beaker for dialysis.



Chemgod1
United States

Send PM
See Mini bio

Status: Frog Egg
Frog Egg
Posted By Chemgod1
on 8/20/2008 11:16 AM   
Reply to this post Go to the top of the page

Thank you all for helping. make's lots of sense.



As a Scientist Solutions member, you are able to register a positive vote for any topic which you believe is useful and relevant to our board or any reply which you believe is especially well worded and helpful.

By participating in the voting, you will be helping to identify the best topics & replies on the board.

You may vote once for any one post, and you may not vote for your own posts.

A post (topic or reply) will earn one "thumbs up" icon for every 10 votes received (up to 3 thumbs up), and the person who made the post will also earn two bonus points.

learn more about member points.



Click here to
Become a member & join our
community (It's easy & free)
Already a member? Please log in
User Name  
Password  
Forget Password?
Find out about our Rewards System
Scientists
Not finding the answer you need?

Post a new topic

You must be logged in to post. Log in above.
Not a member yet? Click here to register
(it's free)
Thank You to Our Sponsor