Hi,
I encountered problem with Abeta 40 aggregation from Inovagen company. We tried many different protocols, pH, but it just wont form any fibrils. Anyone has similar experience? Most often we use alkaline pretreatment similar to this one: https://www.ncbi.nlm.nih.gov/pubmed/17046389, but we skip the lyophilization (we use our peptide directly after treating it with NaOH).
Hi Miroslav. were the ab peptides from Inovagen in DMSO?. In that case you need to remove the DMSO from the peptides by quick gel filtration like PD-10 columns.
No, we dont use DMSO for Abeta 40 fibrils preparation, although we already tried standard Abeta 42 protocol, which includes DMSO and it did not work. Normaly, our steps in Abeta 40 preparation are: resuspension in 10mM NaOH, sonication, centrifugation and dissolvement to 150mM MOPS .. however, this does not work with Inovagen peptide
(we also tried: acidic pH buffer - HCl, neutral pH - PBS, aliqouts prepared from HFIP, aliquots from DMSO)
Hi Miroslav. if these ab40 or ab42 peptides don't dissolve in DMSO even with sonication, I suggest you find another supplier.
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