Hi researchers!
We recently have bought amyloid beta 1-42 peptide from Genscript. According to the material data sheet, the peptide is soluble in water. However, after examining the amino acids, there are close to 45% of hydrophobic amino acids and 28% of charged amino acids.Theoretically speaking, this is not considered to be a hydrophilic peptide.
So, I was looking up methods already recommended by papers to dissolve this peptide. Some recommend dissolving in HFIP to remove the pre-aggregates and dry under a nitrogen stream. Following that, it is recommended to add a small amount of DMSO and the dilute with PBS or water. But some protocols mention that we could directly skip to adding DMSO. I only have 1.0mg of peptide and I am not to confident about adding HFIP and drying it as I might lose some peptides.
Thus, would it be fine to just dissolve in a small amount of DMSO and then dilute further with water? I do not think I can dilute the peptide sample in just water as it does not seem to be hydrophilic. If this method is fine, what would be the recommended volume of DMSO to be added to get a final concentration of 1mg/mL after adding water?
Thanks so much!
-Mathangi
What concentration do you need? Compounds with limited solubility in water still dissolve when their concentration is low enough. I tend to dissolve peptides in a minimum of DMSO, but make sure whatever you will do with the peptide tolerates DMSO.
Use the recommendation from Genscript. Add a buffer at the pH of maximum stability (again, the vendor should provide this) and use a compatible surfactant for biologics at an amount that suits your final API targeted concentration. Be aware that any extreme conditions including organic solvents may generate aggregates.
Jack Silver Thank you!I would need a concentration of 1mg/mL. Alright, so perhaps I can dissolve in a small amount of DMSO (maybe 90uL) and then add a remaining 910uL of water? Would you recommend addition of HFIP? Many protocols and answers by other researchers have suggested this to remove the pre-aggregates.
Mathangi Palanivel I would try the DMSOI as you suggested. I'm not familiar with HFIP, so I'm not really qualified to comment about that. :-)
Hi Dr Mathangi Palanivel . Soluble in DMSO . See the link : https://www.abcam.com/ps/products/120/ab120301/documents/Amyloid%20Beta%20Protocol%20WC050216%20v2%20(website).pdf
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biomolecules
- Peptides