Journal: bioRxiv

Article Title: Ultrasonic Reporter of Kinase Activity

doi: 10.64898/2025.11.30.691048

Figure Lengend Snippet: a) xAM images of UReKA1 and UReKA2 incubated with PKA at various timepoints at 634kPa (for maximal signal). b) Normalized nonlinear US signal of UReKA1 (blue) and UReKA2 (orange) over time (n=3). c) Normalized phosphorylated fraction of UReKA1 (blue) and UReKA2 (orange) over time (n=1). d) Nonlinear ultrasound images (left) and signal (right) of UReKA1 incubated with PKA and phosphorylated UReKA1 incubated with LPP with or without buoyancy purification. (n=2). e-f) Schematics of the proposed molecular mechanism of UReKA with reversible phosphorylation reaction for the phosphorylation as forward reaction (e) and dephosphorylation of GvpC as reverse reaction (f).

Article Snippet: For dephosphorylation assays, 10 units of LPP (P0753L; NEB) per μL of engineered GVs at OD500 = 10 were mixed with PMP Buffer (50 mM HEPES, 100 mM NaCl, 2 mM DTT, 0.01% Brij 35, pH 7.5) and 1 mM MnCl 2 , followed by incubation with slow rotation at 37°C for 10–12 hours.

Techniques: Incubation, Purification, Phospho-proteomics, De-Phosphorylation Assay

Journal: bioRxiv

Article Title: Rare bioactive tau oligomers from Alzheimer brain support both templated misfolding and fibril formation

doi: 10.1101/2025.09.24.678227

Figure Lengend Snippet: A) Schematic representation of HMW tau dephosphorylation by the Lambda Protein Phosphatase (LPP). B) Seeding activity assay using FRET reporter HEK cell; the integrated FRET intensity was quantified by flow cytometry. Data represents mean ± S.D. of three individual experiments performed in triplicate with HMW Tau isolated from the AD case 1971. Results were analyzed using the unpaired student t-test, ** for p < 0.005. C), D) Dot blot quantification of HMW tau, originating from the same case as panel B, treated or not with Lambda PP and probed with the anti-phospho tau AT8 or the total tau antibody D5D8N.

Article Snippet: HMW tau was subjected to dephosphorylation using Lambda Protein Phosphatase (LPP) kit (New England Biolabs (NEB), #P0753S).

Techniques: De-Phosphorylation Assay, Activity Assay, Flow Cytometry, Isolation, Dot Blot