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Dialysis

Separation of substances in solution by means of their unequal diffusion through semipermeable membranes.The process of separating larger molecules from smaller molecules that occurs in kidney dialysis for example, where a machine takes a role of a kidney and filtrates blood.


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Why dialyse DMEM before protein purification (Ni-Affinity)?

... at first sight, I hope all the same it's not too stupid to ask.... It can be reduced to "why is it - in this case - necessary to perform a dialysis?" We transfected HEK293T in order to express a his-tagged protein. The cells were kept in DMEM (high glucose, modified with 5% FCS and ...

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by LittleBird
Sun May 20, 2012 3:43 pm
 
Forum: Molecular Biology
Topic: Why dialyse DMEM before protein purification (Ni-Affinity)?
Replies: 3
Views: 814

HCL instead of imidazole!

... very slowly in several steps. Thus, if you had pH 1 and want pH 8.0, I would do at least eight steps, one pH point per step (i.e. first dialysis against buffer with pH 2, second dialysis with pH 3, third with pH 4...). Does your native protein turn yellow in the reducing buffer. I don't ...

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by JackBean
Fri Dec 16, 2011 11:57 am
 
Forum: General Discussion
Topic: HCL instead of imidazole!
Replies: 6
Views: 1120

HCL instead of imidazole!

... of your column? Do you have protein of which you can measure activity? If so, do it and you will see. You will probably have to do some slow dialysis to re-fold your protein.

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by JackBean
Fri Dec 16, 2011 7:18 am
 
Forum: General Discussion
Topic: HCL instead of imidazole!
Replies: 6
Views: 1120

Hypothetical experiment

... years X = 1,000,000 min = 16,666.67 hrs = 694.44 days = 1.90 years 1. Take some of the E.coli (whole genome is sequenced) and put them into dialysis cup/jar in normal LB broth. 2. Set up three sets of dialysis chambers: a) control – regular LB broth; b) experimental chamber 1: LB broth + ...

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by Cat
Tue Dec 06, 2011 8:54 pm
 
Forum: Evolution
Topic: Hypothetical experiment
Replies: 6
Views: 2251

Denatured protein

that was just idea, since I know, that proteins are sometimes extracted with the use of urea and then refolded during dialysis. But with the acidic environment it should work as well. Slowly increasing pH and the protein could refold.

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by JackBean
Fri Nov 25, 2011 12:44 pm
 
Forum: Molecular Biology
Topic: Denatured protein
Replies: 5
Views: 1217
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