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

... I transfected my units with my develop to prepare my protein in the supernatant. I gathered the supernatant around the range of 1 litre and after dialysis I subjected my supernatant to cleaning yet I don't get my proteins at the closure. Do you have any suggestions?

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by sullivan
Mon Jul 22, 2013 12:28 pm
 
Forum: Molecular Biology
Topic: Why dialyse DMEM before protein purification (Ni-Affinity)?
Replies: 5
Views: 2679

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: 5
Views: 2679

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: 2090

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: 2090

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: 4426
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