We synthesized a peptide made up of 7 amino acids. After cleaving the peptide with TFA, we tried to precipitate the peptide from cold ether. However, it seems like our peptide is somehow soluble in ether. Are there any other organic solvents to precipitate peptides?
To precipitate small peptides which are having a molecular size below 10KDa, Add slowly cold Acetone (-40 degree Celsius) to a volume ratio of 1:200
If your peptide is not precipitating from cold ether after cleavage with trifluoroacetic acid (TFA), there are several other organic solvents you can consider using for peptide precipitation. The choice of solvent depends on the properties of your peptide and the conditions of your experiment. Here are a few alternative organic solvents that you can try:
When choosing an organic solvent for peptide precipitation, consider the following tips:
- Use the solvents at low temperatures, ideally in an ice bath or a freezer, to enhance precipitation.
- Perform a small-scale test with a small amount of your peptide to determine the most effective solvent for your specific peptide sequence and conditions.
- Gentle mixing and avoiding excessive agitation can help prevent peptide denaturation during the precipitation process.
- After precipitation, centrifuge the mixture to separate the solid peptide from the solvent.
Keep in mind that the solubility of peptides can vary based on factors like peptide sequence, side chain interactions, and post-translational modifications. It's important to optimize your conditions to achieve the best results for your specific peptide. If none of the organic solvents seem to work, you might need to consider other purification methods, such as reversed-phase HPLC or solid-phase extraction. Onur Alpturk
You could lyophilize the mixed solution TFA-ether that contains the peptide.
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