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

Journal: Acta Neuropathologica

Article Title: Synaptotoxic effects of extracellular tau are mediated by its microtubule-binding region

doi: 10.1007/s00401-025-02897-0

Figure Lengend Snippet: MTBR/R’ is required for acute plasticity disruption by synaptotoxic tau in AD brain extracts. a Aqueous extracts from the brains of two people with AD (AD1 and AD6) were analysed by Western blotting in the presence or absence of lambda protein phosphatase (± LPP) using the monoclonal anti-tau antibodies Gen2A and Gen2B. Migration of SDS-PAGE molecular weight standards (in kDa) is indicated on the left. The bracket on the right highlights the position of full-length tau whereas arrows indicate likely tau fragments detected by both mAbs. It should be noted that AD6 extract had much higher total tau concentration as measured by immunoassay . b Unlike isotype control IgG, anaesthetised rats co-injected with AD1 extract (10 µL) and either Gen2A or Gen2B antibodies (2.5 µg i.c.v.) maintained normal hippocampal LTP. c Similarly, robust LTP was induced when Gen2B mAb (2.5 µg i.c.v.) was co-administered with the extract AD6 (10 µL). Left-hand panels in b, c show the time course of LTP. Summary bar charts of LTP magnitude during the last 10 min are displayed in the right-hand panels. Values are mean ± SEM. ## p < 0.01, ### p < 0.001 compared with pre-HFS, paired t -test; *** p < 0.001, **** p < 0.0001, one-way ANOVA followed by Bonferroni’s multiple-comparison tests in b and unpaired t -test in c

Article Snippet: Aqueous extracts from the brains of two people with AD (AD1 and AD6) were analysed for tau protein by western immunoblotting in the presence or absence of lambda protein phosphatase (LPP) concentrate supplied at 400,000 units/ml, diluted 1:50 to 8000 units/ml for final reaction as per kit recommendation (New England Biolabs P0753S).

Techniques: Disruption, Western Blot, Migration, SDS Page, Molecular Weight, Concentration Assay, Control, Injection, Comparison