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Questions tagged [chromatography]

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2
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0answers
23 views

Can you convert smells to variables?

Is there a way to map smells to numerical values of different variables? I.e. With RGB colours, each colour is made up of numerical values from 0-255. For example, red is (255,0,0), and yellow is (...
2
votes
2answers
47 views

How can I describe running a solution four times through a chromatography unit (in an instruction)?

I'm translating a document from Russian: The mean retention time for the first four injections of RNase should be (40.8± 2) min. If the mean RT is beyond this range, adjust the gradient system (...
2
votes
2answers
362 views

Ion-exchange vs Gel Permeation Chromatography for proteins

For extracting Lysozyme from egg white which method would you recommend and please help me understand why. Lysozyme has a higher isoelectric point (pH 10.5) and a lower molecular mass (14 307 Da) ...
3
votes
1answer
108 views

Sephadex column for alpha-amylase

I want to purify crude alpha-amylase with column chromatography. I am using a spehadex 75, But for some reason I can't find any information on how to make the slurry. I can quickly find tons of ...
5
votes
2answers
1k views

What are “primary amino acids and secondary amino acids”? Context: analysis of amino acid content using reversed-phase HPLC

In a Russian document I'm translating, an HPLC system is used to analyse the amino acid content of a substance. The detector wavelength is set at 262 nm for "secondary amino acids" and 338 nm ...
7
votes
2answers
171 views

Is there a term for a procedure in which the chromatography column is washed with 20% alcohol

From a method description in a Russian document: After the chromatographic analysis is complete, the column is flushed with at least 2-3 volumes of water at a rate of 0.4 ml/min. The column is then ...
3
votes
1answer
1k views

Clarify pre-column pressure vs. system pressure vs. backpressure for prepacked columns used with the äkta purification system

Background: I am using the Äkta pure system for protein purification, with prepacked columns from GE-healthcare. Just to take one example, I use the HisTrap 5 mL column (IMAC: immobilized metal ...
0
votes
1answer
1k views

What is DNA-cellulose chromatography?

I am going through a paper where they have done DNA-cellulose chromatography to study the interaction between protein and DNA. They are adding the protein to the column and eluting with high salt ...
0
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0answers
12 views

Hydrophobic elution times in ionic exchange columns

What would the order of elution be of say Gly, Val, Leu? My argument is that hydrophobic residues will try and get away from the resin and the more hydrophobic the residue is the faster it will elute....
2
votes
2answers
29 views

Affinity Column of membrane bound receptors

I understand affinity columns can be used to study the ligand/enzyme affinity. But is an affinity column able to be used for membrane bound receptors? I
2
votes
2answers
1k views

View ABI chromatogram plots with python

I would like to view the chromatogram traces from a few ABI (.ab1) files. I would prefer to use python for this, or a function with python bindings, or at least some open source package such as EMBOSS....
3
votes
1answer
80 views

Decreasing Solubility as Solvent Moves Higher in Paper Chromatography

In paper chromatography, as the mobile phase (I used 1:9 acetone/petroleum ether) climbs higher and higher, will the solubility of the solutes decrease as the solvent moves higher? I speak mostly ...
6
votes
2answers
703 views

Ni-NTA purification, problem with the chaperone protein

I'm trying to purify the protein by Ni-NTA affinity chromatography. It seems that my protein (size about 54 kDa) is co-purified with chaperone protein (probably GroEL - as the band is around 57kDa). ...
4
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2answers
3k views

Why digest proteins into peptides for Liquid Chromatography - Mass Spectrometry?

Digesting (trypsin or whatever other proteolytic enzyme) proteins generates multiple peptides so the degree of complexity of the sample, at the peptide level, increases a lot. In addition there is ...
2
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1answer
1k views

How does DNA resolve on size exclusion resin?

We generally have a good idea of how DNA separates using agarose gel electrophoresis, how well does DNA resolve on a SEC resin like superose? I get the impression that salt influences where it elutes. ...