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Sepharose

Definition/Description

A registered trademark owned by GE Healthcare companies for the bead-formed gel prepared from agarose, which is used as base matrix for several chromatographic products used for purification of proteins and other biomolecules.


Supplement

In its natural state agarose it occurs as part of the complex mixture of charged and neutral polysaccharides referred to as agar. The agarose used to make SepharoseTM is obtained by a purification process which removes the charged polysaccharides to give a gel with only a very small number of residual charged groups.

SepharoseTM is given different chromatographic properties by modification or attachment of a ligand.


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Lost Protein?

... column is 20mM TRIS(pH 8.5)... and I salt out using KCl across the concentration gradient of 0.1M - 1.0M... and the column is a anion exchanger, sepharose (Q). Now the problem is I'm not seeing my protein after elution, roughly about 20 times the volume of the column has been passed through (again, ...

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by BrianScott
Tue Jun 30, 2009 4:05 pm
 
Forum: Molecular Biology
Topic: Lost Protein?
Replies: 0
Views: 143

beta lactamase

Sorry. at pH 7-7.5 lactamase from E. coli should be postively charged and bind to S-sepharose, I meant to say. It should *not* bind to Q-sepharose.

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by blcr11
Sat Mar 31, 2007 8:28 am
 
Forum: Microbiology
Topic: beta lactamase
Replies: 2
Views: 1091

beta lactamase

... by chromatography. I can’t say for sure, since I don’t know the specifics of the lactamase you’re trying to purify, but I don’t think Q-sepharose is the resin of choice for you. It depends on the pI of your protein. Penicillinase (bla) from E. coli has a theoretical pI of 9.4, according ...

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by blcr11
Sat Mar 31, 2007 8:23 am
 
Forum: Microbiology
Topic: beta lactamase
Replies: 2
Views: 1091

beta lactamase

hi please i want to purify the enzyme of beta lactamase using Q sepharose fast flow.i haven't any idea.can you help me please?have you an interest articals?? thanks

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by naouel_24
Tue Mar 13, 2007 8:58 pm
 
Forum: Microbiology
Topic: beta lactamase
Replies: 2
Views: 1091


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