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In contrast to amino acids, growth factors activate mTORC1 by stimulating the dissociation of the tuberous sclerosis complex (TSC) from the lysosome and Rheb (Reference Menon, Dibble and Talbott83)

Kit Components Additional Materials Required Microplate reader capable of measuring absorbance at 340 nm at 25C Precision pipettes to deliver 2 L to 1 mL volumes Multi-channel pipettes to deliver 20 L to 200 L volumes Tubes to prepare sample dilutions 15 mL conical tubes Incubator at 25C Deionized water Assay Procedure Summary Prepare Working Solution

Calvert JW, Jha S, Gundewar S, Elrod JW, Ramachandran A, Pattillo CB, et al

M., Yahya, A

Proteins were eluted by heating in SDS loading buffer at 90 C for 5 min, and lipoylation phenotypes were verified by immunoblotting the eluate alongside the input control