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EDT extraction and isolation from peptide cleavage mixture

EDT extraction and isolation from peptide cleavage mixture - Peptide Forum

EDT extraction and isolation from peptide cleavage mixture - Peptide Forum. Ask and discuss questions on peptide protocols, custom peptide synthesis, peptide identification and peptide sequencing.


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  #1  
Old 09-22-2006, 06:38 PM
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Default EDT extraction and isolation from peptide cleavage mixture



Hi, I'm trying to isolate EDT effectively without stenching up my whole lab with toxic fumes. I've been using an ether precipitation/centrifugation method. This is quite tedious, as it entails multiple ether precipitation/centrifugations and my centrifuge isn't in the hood which isn't good and I'd rather choose another method. I guess I could try to filter my solution, but it seems I lose a lot of my peptide due to the filtering. I'm thinking of getting a roto-vap in the hood, any good pros and cons to this? Or would getting a spark-free centrifuge be sufficient, inside the hood?
Also, I have a peptide in aqueous PBS buffer now that still has EDT contamination. Any tips on how to get rid of the EDT besides lyophilization? I tried a liquid-liquid extraction 1:1 with Ether but I didn't see any layers at all.

Thanks in advance!

peptidenoob
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Old 09-25-2006, 08:34 AM
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Default Re: EDT extraction and isolation from peptide cleavage mixture

How concentrated is your peptide? Is HPLC a good method for this separation?
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Old 09-25-2006, 06:31 PM
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Default Re: EDT extraction and isolation from peptide cleavage mixture

I have about 100 mgs of this stuff. We were thinking of HPLC, but the stench is bad enough where we might not be able to do it in the lab. I was thinking of maybe a pH precipitation, and then spinning it down...but not sure if this would work or not....
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Old 09-26-2006, 05:25 PM
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Default Re: EDT extraction and isolation from peptide cleavage mixture

Hey peptidenoob,
welcome to the forum!

Can one use protein precipitation methods? you may get away with precipitating your peptides and then reconstituting them into another buffer.

Although your peptides are quite precious and expensive, I would look into this before attempting.
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Old 09-26-2006, 07:43 PM
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Default Re: EDT extraction and isolation from peptide cleavage mixture

yes...I agree thanks
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Old 08-14-2008, 09:11 AM
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Default Re: EDT extraction and isolation from peptide cleavage mixture

Try precipitation in ehter:n-pentane 1:1. After centrifugation remove the ether/pentane layer and let the peptide dry for 1.5 hour. Disolve it in H2O: ACN 1:1 and then freezedry it for 36 hours.
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