Datasheet

生物学的記述

Specificity	Phospho-MBS/MYPT1 (Thr696) Antibody [K5H5] detects endogenous levels of total MBS/MYPT1 protein only when it is phosphorylated at Thr696.
Background	Phospho-MBS/MYPT1 (Thr696) marks a critical phosphorylation site on the myosin-binding subunit (MYPT1) of the myosin phosphatase complex, which pairs the PP1 catalytic subunit with MYPT1 and M20 to precisely dephosphorylate myosin light chain (MLC), thereby fine-tuning actomyosin contractility across diverse cell types. MYPT1 contains leucine-rich repeats that anchor PP1c and inhibitory phosphorylation motifs, placing Thr696 within a flexible region highly susceptible to kinase action. RhoA activation triggers ROCK to phosphorylate Thr696, prompting a conformational change that occludes the PP1c active site and sharply reduces its ability to dephosphorylate MLC, allowing phosphorylated MLC to persist and drive robust cytoskeletal tension. This mechanism feeds into the Rho/ROCK/MLC phosphatase axis, where Thr696 modification synergizes with CPI-17 phosphorylation to amplify signaling outputs, sustaining actomyosin interactions even under subsaturating calcium conditions. The result cascades into enhanced stress fiber assembly, focal adhesion reinforcement, and dynamic cell shape changes essential for migration, cytokinesis, and tissue morphogenesis in skeletal muscle and non-muscle cells. In skeletal muscle fibers, this phosphorylation modulates force generation and sarcomere organization during contraction-relaxation cycles, influencing motility and repair processes without relying on vascular-specific tone. Such dysregulation, often from excessive ROCK activity, contributes to fibrotic stiffening and impaired regeneration in musculoskeletal disorders.

Specificity

Phospho-MBS/MYPT1 (Thr696) Antibody [K5H5] detects endogenous levels of total MBS/MYPT1 protein only when it is phosphorylated at Thr696.

Background

Phospho-MBS/MYPT1 (Thr696) marks a critical phosphorylation site on the myosin-binding subunit (MYPT1) of the myosin phosphatase complex, which pairs the PP1 catalytic subunit with MYPT1 and M20 to precisely dephosphorylate myosin light chain (MLC), thereby fine-tuning actomyosin contractility across diverse cell types. MYPT1 contains leucine-rich repeats that anchor PP1c and inhibitory phosphorylation motifs, placing Thr696 within a flexible region highly susceptible to kinase action. RhoA activation triggers ROCK to phosphorylate Thr696, prompting a conformational change that occludes the PP1c active site and sharply reduces its ability to dephosphorylate MLC, allowing phosphorylated MLC to persist and drive robust cytoskeletal tension. This mechanism feeds into the Rho/ROCK/MLC phosphatase axis, where Thr696 modification synergizes with CPI-17 phosphorylation to amplify signaling outputs, sustaining actomyosin interactions even under subsaturating calcium conditions. The result cascades into enhanced stress fiber assembly, focal adhesion reinforcement, and dynamic cell shape changes essential for migration, cytokinesis, and tissue morphogenesis in skeletal muscle and non-muscle cells. In skeletal muscle fibers, this phosphorylation modulates force generation and sarcomere organization during contraction-relaxation cycles, influencing motility and repair processes without relying on vascular-specific tone. Such dysregulation, often from excessive ROCK activity, contributes to fibrotic stiffening and impaired regeneration in musculoskeletal disorders.

使用情報

Application

ELISA, IF, WB

Dilution

WB

1:1000-1:2000

Reactivity

Chicken, Human, Mouse, Rat

Source

Mouse Monoclonal Antibody

MW

115 kDa

Storage Buffer

PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage
(from the date of receipt)

-20°C (avoid freeze-thaw cycles), 2 years

プロトコール

References

https://pubmed.ncbi.nlm.nih.gov/12600888/
https://pubmed.ncbi.nlm.nih.gov/24712327/

Application Data

WB

Validated by Selleck

Lane 1: Jurkat (serum-starvation overnight), Lane 2: Jurkat (serum-starvation overnight; Calyculin A, 100 nM, 37℃, 30 min)