atpase elipa kit  (Cytoskeleton Inc)

Journal: bioRxiv

Article Title: N-terminal phosphorylation inhibits Arabidopsis katanin and affects vegetative and reproductive development in opposite ways

doi: 10.64898/2026.01.20.700631

Figure Lengend Snippet: DDD has lower microtubule binding affinity and ATPase activity in vitro . (A) Representative Coomassie Blue-stained SDS-PAGE gels of microtubule co-sedimentation assays. 1 µM KTN1 (n = 3), DDD (n = 3), or AAA (n = 6) was incubated with the indicated concentration of taxol-stabilized microtubules. S, supernatant; P, pellet. Arrows identify the different protein bands. (B) Binding curves of KTN1, DDD, and AAA proteins corresponding to (A) . Each data point represents the mean ± SEM. Data were fit to a one-site saturation binding model to obtain the microtubule binding affinity and maximum amount of protein binding. (C-E) Plots of ATPase activity over time of 25 nM of KTN1 (C) , DDD (D) , and AAA (E) proteins. ATPase activity was measured as the amount of Pi release in the absence (black) or presence of either 25 nM (blue) or 250 nM microtubules (red). Each data point represents the mean ± SEM from 9 independent experiments.

Article Snippet: The ATPase activity of 25 nM of KTN1, DDD, or AAA in the presence of either 25nM or 250nM of taxol-stabilized microtubules was evaluated using a commercially available ATPase ELIPA Kit (Cytoskeleton #BK051) according to the manufacturer’s instructions.

Techniques: Binding Assay, Activity Assay, In Vitro, Staining, SDS Page, Sedimentation, Incubation, Concentration Assay, Protein Binding

Journal: Saudi Pharmaceutical Journal : SPJ

Article Title: Design, synthesis, molecular docking, and in vitro studies of 2-mercaptoquinazolin-4(3 H )-ones as potential anti-breast cancer agents

doi: 10.1016/j.jsps.2024.101971

Figure Lengend Snippet: Inhibition assay of the synthesized quinazolinone compounds (3a, 3b, 3e, 3 g, and 3 h) against KSP (% inhibition at 2 µM and IC 50 ), using Ispinesib as positive control, and against PI3Kδ using Idelalisib as positive control.

Article Snippet: For IC 50 value measurement, a microtubule-activated ATPase kinetic assay kit (Cytoskeleton, Cat. # BK060) was used as per the manufacturer’s instructions ( ).

Techniques: Inhibition, Synthesized, Positive Control, Control, Activity Assay

Journal: Biomedicines

Article Title: Interaction of Tau with Kinesin-1: Effect of Kinesin-1 Heavy Chain Elimination on Autophagy-Mediated Mutant Tau Degradation

doi: 10.3390/biomedicines12010005

Figure Lengend Snippet: Kinesin-1 motor domain interacts with tau in vivo and in vitro. ( A ). Diagrammatic representation of KIF5B fragments and the motor domain of KIF5B is indicated within the red frame. ( B ). Western blot of GST-tagged KIF5B fragments (motor + stalk, neck and tail region) with pull down of tau from P301S tau mouse brain lysate and the brain lysate was loaded as input. The purity of KIF5B fragments was ascertained by Commassie blue staining of SDS -PAGE gel (lower panel). ( C ). Western blot of GST-KHC-Motor with pull down of tau from P301S tau mouse brain lysate. ( D ). Western blot of GST-KHC-Motor with pull down of recombinant 2N4R tau protein. The tau levels in the precipitates (ppt) and supernatants (sup) of samples loaded with GST-Kinesin motor coupled beads or beads alone were demonstrated.

Article Snippet: Kinetic measurements of microtubule (MT)-activated kinesin ATPase activity, with or without tau, were performed using a Kinesin Enzyme Linked Inorganic Phosphate Assay (ELIPA) kit (Cytoskeleton, Inc., Denver) as per the manufacturer’s instructions and as described elsewhere [ , ].

Techniques: In Vivo, In Vitro, Western Blot, Staining, SDS Page, Recombinant

Journal: Biomedicines

Article Title: Interaction of Tau with Kinesin-1: Effect of Kinesin-1 Heavy Chain Elimination on Autophagy-Mediated Mutant Tau Degradation

doi: 10.3390/biomedicines12010005

Figure Lengend Snippet: Regulation of the motor activity of KHC-Motor by recombinant tau 441 2N4R ( A ). The ATPase activity of the KHC-motor (50 nM) is stimulated by microtubules (MT; 300 nM) in ELIPA assay, which was performed with or without different concentrations of recombinant tau (1.25 to 10 μM). ( B ). The increasing concentrations of tau 2N4R inhibit the ATPase activity of the kinesin motor from 0 to 5 min. The data were presented as the mean± SEM from three separate experiments. **** p < 0.0001 vs. KHC-Motor alone.

Article Snippet: Kinetic measurements of microtubule (MT)-activated kinesin ATPase activity, with or without tau, were performed using a Kinesin Enzyme Linked Inorganic Phosphate Assay (ELIPA) kit (Cytoskeleton, Inc., Denver) as per the manufacturer’s instructions and as described elsewhere [ , ].

Techniques: Activity Assay, Recombinant