I am assaying the effect of an activator peptide on degrading monomeric beta amyloid. After effecting my peptide on extract cell and a complex of monomer and oligomers and fibril of beta amyloid, I must detect remained monomeric amyloid beta.
How can I detect only remained monomeric beta amyloid after using my peptide?
please help and guide me.
Probably the unique way is using Mass spectrometry but if cell extract must be also present it would be also difficult to look only for abeta forms. I wonder whether you may want to detect just the remaining monomer or the products of degradation.
Dear Franco
Thanks for your helping. Please answer this question:
I want use HFIP for aliquoting 0.5 mg to several episode for next uses. but I do not know how much HFIP I must use. Please guide me.
Thanks
Perfect. You have two options. One is the most standard for peptides, which is to prepare a concentrated solution in DMSO and do aliquots. If you decide to use this approach you may prepare a 1 mM solution adding (to the 0.5 mg) 115 micro liters of DMSO. And then aliquot in 5-10 micro liters and store them at -20 or -80 degrees. When you have to use aliquots ,you take one aliquot from the freezer, wait for thawing, pipette the suitable amount of peptide solution and dilute it in medium or PBS.
Second option is similar but with the reagent you mentioned: HFIP.
In DMSO and storing aliquots in the freezer you would not expect much change upon time. With HFIP I am not so sure what would happen frozen although if stored unfrozen there is some changes in structure (as reported in different papers). I would see details in this paper that is attached.
Good luck
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biomolecules
- Peptides