I am doing tht test for assaying beta amyloid aggregation. I had two sample, one of them was "amyloid+ tht" as a positive control and the another was "my extract cell + amyloid+tht" and then we use 440 and 480 as excitaion and emission wavelenght.
But I saw that measured aggregation of beta amyloid at present cell extract is higher than the absent of cell extract.
In the other word, I want to know
1- why cell extract increases the beta amyloid aggregation?
2- Is it logical?
Please help me.
Thanks
First of all you should make a control consisting of cell extract plus Tht. Cell extract is a very complex mixture of molecules that can bind Tht and change is emission spectrum. This "blank" should be subtracted from your experimental sample.
I agree with the suggestion given by Stefania Rigacci. Its true that cell extract is complex mixture and it is possible that any of present molecule show synergistic effect, giving more augmented ThT binding than with beta amyloid alone. The set of condition you are following will give you ambiguous result, so it would be better to go for "Blank"
Hi,
Cell membranes tend to accelerate amyloid formation, it is quite a common feature of this family of proteins, and especially the ones present in neurons. I refer you to this recent article that I enjoyed reading, regarding a-synuclein and lipid vesicles (Lipid vesicles trigger α-synuclein aggregation by stimulating primary nucleation) :
http://www.nature.com/nchembio/journal/v11/n3/abs/nchembio.1750.html
Benjamin
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