5 votes
Accepted

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

Prepared or conditioned for storage would probably be perfectly acceptable. If you're looking for a single-word equivalent, in this case, I would translate as stabilized. From dictionary.com: ...
hBy2Py's user avatar
  • 166
5 votes

How can I change my buffer system for protein purification?

There are a number of ways to address this, and the other answers are certainly correct. Another strategy, especially if you already have the sample in PBS, is just to dilute it in the low salt ion ...
canadianer's user avatar
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5 votes

How can I change my buffer system for protein purification?

Assuming that your protein can handle being in the new buffer, you have multiple options. Dialysis will work although you might want to consider using a centrifuge spin filter with a suitable MW ...
Jeppe Nielsen's user avatar
4 votes

Question in gel chromatography experiment

Within reason, you can basically use any buffer you like with Sephacryl S-100, provided that is suits your purpose in terms of protein/enzyme stability, and phosphate-buffered saline sounds ideal. ...
user338907's user avatar
  • 4,678
3 votes

Sephadex column for alpha-amylase

The '75' of Sephadex G-75 refers to the water regain value, which is defined as the grams of water absorbed upon hydration of 1g dry powder, multiplied by 10 (see Reiland). Thus the water-regain value ...
user338907's user avatar
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3 votes
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What are "primary amino acids and secondary amino acids"? Context: analysis of amino acid content using reversed-phase HPLC

I would guess that 'secondary' refers to amino acids that have been derivatized in some way, possibly by reacting with o-phthalaldehyde. (To measure the fluorescence of an OPA-derivatized amino acid, ...
user338907's user avatar
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3 votes

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

I would continue to use inject to describe loading samples into the liquid chromatography system and replace chromatograph the RNAase solution four times. with ...
Michael_A's user avatar
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3 votes
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How to produce and isolate an enzyme?

Chromatography is one of the best ways to isolate a protein, full stop. It is extremely widely used in both academia and industry. Before we get into the details, though, a definition is in order. ...
MattDMo's user avatar
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3 votes

How can I change my buffer system for protein purification?

Two accepted methods of 'desalting' a protein and/or changing the buffer are dialysis and gel-filtration. Dialysis is time-consuming, and gel-filtration usually requires a concentrated sample (It is ...
user338907's user avatar
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2 votes
Accepted

View ABI chromatogram plots with python

The Biopython 1.65 ABI parser should expose the chromatogram data, as of Biopython 1.66 it should expose everything. UPDATE: Example using this for plotting here: http://biopython.org/wiki/ABI_traces
Peter Cock's user avatar
2 votes

How can I change my buffer system for protein purification?

Impossible to answer this in detail without knowing why you started with PBS, but simpler is better so if you can start with a buffer that is compatible with the ion exchange step then I would try ...
tyersome's user avatar
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2 votes

What buffer should I choose for IEX chromatography for purifying IgY

Your column manual will probably come with buffer recommendations (these manuals are also often available online). You can also search for publications that have purified IgY by anion exchange ...
canadianer's user avatar
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2 votes
Accepted

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

I would say simply: repeat the chromatography four times or less formal, but quite acceptable: re-run the column (or whatever it is) four times This puts the emphasis on the process. But I’m ...
David's user avatar
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2 votes

Ion-exchange vs Gel Permeation Chromatography for proteins

Here is what I have come up with. Thank you for all your help. Ion Exchange Chromatography(IEC) will yield the purist product from a single application. The main reason is due to the fact that Gel ...
Cello_101's user avatar
2 votes
Accepted

Ion-exchange vs Gel Permeation Chromatography for proteins

Since lysozyme is so well studied, your best bet is to search the literature and see what methods other people use. See this paper, for example. They report that lysozyme can be affinity purified on ...
canadianer's user avatar
  • 17.6k
2 votes

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

This is a routine procedure in FPLC to prepare columns for storage. 20% ethanol is used to prevent microbial growth. See this technical note: All SEC columns are delivered in 20% ethanol to prevent ...
canadianer's user avatar
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2 votes
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Clarify pre-column pressure vs. system pressure vs. backpressure for prepacked columns used with the äkta purification system

Is there a good article or webpage that goes through this theory at a basic level? Yes, Chapter 7 of the manual linked to by Andreas Tosstorff is a good source, and is also the source of the images ...
canadianer's user avatar
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2 votes
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What is DNA-cellulose chromatography?

DNA-cellulose chromatography is used to purify proteins that bind DNA (Potuzak and Dean 1978, Abdullah et al., 1985). The idea behind it is quite simple: you attach DNA to a cellulose matrix and keep ...
AlexDeLarge's user avatar
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1 vote

Affinity Column of membrane bound receptors

This is a classic reference where Stanley Cohen and Graham Carpenter used an affinity column made out of the epidermal growth factor (EGF), and used it to purify the EGF receptor: JBC 1980
mdperry's user avatar
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