Dear Debanjan

what di you mean for real time?

did you mean "in vivo" (eg mouse model) or "in vitro" ?

if you would like perform in vitro studies, i think that you can certanilly monitor the kinetics of fibril formation using the Thiot assay as we have done several year ago for apo-sod1 Wt and mutants is als.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1899188/ (fig. 1)

i'm not an expert of lysozime amyloids but it seems that this assay work also for this fibrills https://www.ncbi.nlm.nih.gov/m/pubmed/23280648/

good luck

ciao

Manuele

Dynamic light scattering would be a very sensitive method for following protein aggregation over time. The equipment is easy to use and does not require large amounts of material.

Hi Debanjan,

you can use Thioflavin T (ThT) dye, it is an amyloid specific probe, using fluorescence spectrophotometer or plate reader you can in real time follow the amyloid aggregation kinetics. Increased ThT intensity you will observe if lysozyme is forming amyloid type of aggregation and in the case of inhibition study decreased ThT intensity compared to control you should observe. you can follow my article for various biophysical tehniques to follow kinetics of both aggregation and inhibition.

Article Structural perturbation by arsenic triggers the aggregation ...

To Adam's point of using DLS, the model will be important. Here are a few references my former labmate used:

Article Monitoring protein assembly using quasielastic light scatter...

Article Assembly of Amyloid Protofibrils via Critical Oligomers—A No...

as well as his paper, though his model may be overkill

Article β-Lactoglobulin Assembles into Amyloid through Sequential Ag...

I followed alpha-synuclein amyloid formation with Thioflavin T similar to Manuele and Neelakant. Some researchers worry about formation of singlet oxygen with the excitation of the ThT. I'm agnostic on the point (maybe there's a definitive paper now?), but if you can degas your solvent you might sleep a little better.

Article In Vitro Formation of Amyloid from ??-Synuclein Is Dominated...

I am highly give obliged to one and all who took their time out to give my question a read and replied so diligently and also for sharing various papers and information which will help me in designing my experiments. I hope for a fruitful conversation on research in future. My best wishes to everyone!:)