Phospho-MBS/MYPT1 (Thr696) Antibody [K5H5]

Application: Reactivity: Chicken, Human, Mouse, Rat

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

Usage Information

Dilution
WB1:1000-1:2000

Application
ELISA, IF, WB

Reactivity
Chicken, Human, Mouse, Rat

Source
Mouse Monoclonal Antibody

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

Predicted MW Observed MW
115 kDa 135 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Datasheet & SDS

Biological Description

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

Clone
K5H5

Synonym(s)
MBS; MGC133042; MYPT1

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.

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.

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

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%