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Gradient elution

Gradient elution

elution in column chromatography in which a changing ph or ionic strength is used to separate substances.

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Wrong imidazole conc

... elute your protein so you do not need to add more, and you can use your elution to do all your experiments further. NAd next time you can use any ... am right it appears that you were going through the motions of doing a gradient elution on you column (Ni-NTA I presume) but were not checking for ...

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by canalon
Mon Oct 31, 2011 10:04 am
Forum: Molecular Biology
Topic: Wrong imidazole conc
Replies: 3
Views: 2031

Biochemistry/ Proteins & Amino Acids

... c) 18 salt bridges d) 20 salt bridges e) 30 salt bridges 2 of 3 6. The elution profiles for the first three cycles of Edman chemistry in the protein ... allows the free amino acid to be separated and identified using a salt gradient on an ion exchange column. 11. _false_____ Of the negative allosteric ...

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by SimplyMe
Tue Feb 23, 2010 8:32 am
Forum: General Discussion
Topic: Biochemistry/ Proteins & Amino Acids
Replies: 4
Views: 6540

Lost Protein?

... 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 ...

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by BrianScott
Tue Jun 30, 2009 4:05 pm
Forum: Molecular Biology
Topic: Lost Protein?
Replies: 0
Views: 2227


If you're using an fplc instrument, you're better off using a gradient of imidazole for elution. Then your stuff will elute at whatever minimal concentration of imidazole that suits ...

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by blcr11
Thu Feb 14, 2008 12:09 pm
Forum: Molecular Biology
Topic: Ni-NTA-Columns
Replies: 3
Views: 2441

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