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Affinity column

Affinity column --> affinity chromatography

(Science: investigation) a technique of analytical chemistry used to separate and purify a biological molecule from a mixture, based on the attraction of the molecule of interest to a particular ligand which has been previously attached to a solid, inert substance.

The mixture is passed through a column containing the ligand attached to the stationary substance, so that the molecule of interest stays within the column while the rest of the mixture continues through to the end. Then, a different chemical is flushed through the column to detach the molecule from the ligand and bring it out separately from the rest of the mixture.


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G genes

... G genes - Recombinant His-tagged proteins (and how they are purified by affinity chromatography on a nickel-agarose column) Thank you very much for your help! - Tim

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by zhetimxu
Sun Jun 28, 2009 7:21 pm
 
Forum: General Discussion
Topic: G genes
Replies: 1
Views: 59

binding on the Affinity column

I HAVE BEEN TRYING TO PURIFY MY GST-FUSION PROTEIN USING THE GLTATHIONE AGAROSE COLUMN AND AFTER RUNNING MY FRACTION SAMPLES ON SDS PAGE I ALWAYS SEE THREE BANDS INSTEAD OF ONE BAND ON MY ELUTED FRACTION. CAN YOU PLEASE HELP ME SO AS TO PURIFY MY FUSION PROTEIN ...

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by VeeKAY
Thu Jul 10, 2008 10:39 am
 
Forum: Molecular Biology
Topic: Help with mRNA purification and DNA polymerase!
Replies: 8
Views: 1806

beta lactamase

... A typical first-time gradient is 0-1M NaCl applied over a 10-20 column volume; you can fine tune the gradient once you know where your protein ... the protein, and make a crude extract (I’m assuming you’re not using an affinity-tagged protein) in no or low salt buffer at pH 7-7.5. (If your protein ...

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

Elimination of low-range co-purified proteins

If you're running a SDS-PAGE column why are your smaller proteins even still around? They should be long gone by the time a 100kDa fraction comes out. Honestly, I'd start over and put an affinity tag on the protein and use affinity columns.

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by Jelanen
Wed Mar 14, 2007 11:43 pm
 
Forum: Molecular Biology
Topic: Elimination of low-range co-purified proteins
Replies: 3
Views: 690

The Fiber Disease

... changes....even the line up and formation of my history column off to the side. Now, If I'm doing something wrong, by all means and ... As substitute antibodies phages demonstrate many features (high affinity, field stability, low cost etc.), which allow considering them as ...

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by London
Mon Dec 11, 2006 8:19 pm
 
Forum: Human Biology
Topic: The Fiber Disease
Replies: 7403
Views: 748960
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