I want to produce amyloid fibril of Abeta1-42 peptide in pH 6 medium ? Is there any specific protocol for this ? or simply diluting the Abeta42 peptide with desired concentration into pH 6 buffer and incubating the peptide at 37 degree Celsius will produce the fibrils ?
Can anyone provide suggestions regarding this aspect ?
Any pH from 5.8 to 8.5 can be used for amyloid fibril formation. therefore pH 6 should not be a problem. You may go on with the method you are using
Generally you will want to employ a monomerisation step, where you break up any possible oligomers/pre-fibrillar species in your powder before self-assembly. This can vary on the peptide in question, though often it involves solubilising the peptide at a high concentration in HFIP or NaOH and sonicating. Then the HFIP is either evaporated off, or the HFIP/NaOH can simply be diluted. Other groups sometimes perform size-exclusion chromatography but unless you are working with a large amount of peptide you will probably lose too much for this to be a viable option, as the peptide will stick to the column.
Otherwise you should be fine simply solubilising your peptide with your buffer at the desired pH. At higher concentrations, you may need to add a small amount of DMSO to adequately dissolve it.
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