Analysis Note

ControlHEK293 cell lysate phosphorylated by ROKα/ROCKII.

Application

Anti-phospho-MYPT1 (Thr850) Antibody, detects level of phospho-MYPT1 (Thr850) & has been published & validated for use in WB.

General description

Phosphorylation of myosin is crucial to smooth muscle contraction. The degree of myosin phosphorylation is dependent in part by the activity of myosin phosphatase. Recent findings suggest that this enzyme can be regulated, which has lead to interest in characterizing this phosphatase. Myosin phosphatase seems to be comprised of three subunits (catalytic subunit of type 1 phosphatase-δ (PP1) and two non-catalytic subunits, large and small (M20)). Studies suggest that the large subunit is a targeting subunit and is termed myosin phosphatase target subunit (MYPT). MYPT has been shown to be a substrate for the Rho-associated kinase (ROK). ROK mediates an agonist-induced increase in myosin phosphorylation and force by inhibiting myosin light chain phosphatase activity through phosphorylation of MYPT.

Immunogen

Epitope: pThr850(Ch) or pThr853(H)

KLH-conjugated, synthetic peptide containing the sequence R30;RS[pT]GVR30;, in which [pT] corresponds to phosphorylated threonine at amino acid residue 850 of chicken myosin phosphatase target subunit 1 (MYPT1). Corresponds to Thr853 in human MYPT1.

Linkage

Replaces: 05-773

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Quality

Westerin Blot Analysis: A 1:500-1:1000 dilution of this antibody detected MYPT1 in RIPA lysates from HEK293 cells phosphorylated by preincubation with ROKα/ROCK-II, active (Catalog # 14-338).

Specificity

Predicted to cross-react with mouse and rat.

Recognizes MYPT1 phosphorylated on Thr850, doublet at Mr ~130 kDa. A non-specific protein was also detected, Mr ~55-60 kDa.

Target description

Doublet at ~130 kDa