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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, PIA | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | ABS45 |
| Replaces | 07-251 |
| Description | Anti-phospho-MYPT1 (Thr696) Antibody |
| Alternate Names |
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| Background Information | Protein phosphatase 1 regulatory subunit 12A (UniProt: O14974; also known as Myosin phosphatase-targeting subunit 1, Myosin phosphatase target subunit 1, Protein phosphatase myosin-binding subunit) is encoded by the PPP1R12A (also known as MBS, MYPT1) gene (Gene ID: 4659) in human. Myosin light chain phosphatase with its regulatory subunit, myosin phosphatase target subunit 1 (MYPT1), modulates Ca2+-dependent phosphorylation of myosin light chain by myosin light chain kinase, which is essential for smooth muscle contraction. Myosin phosphatase regulates the interaction of actin and myosin downstream of the guanosine triphosphatase Rho, which inhibits myosin phosphatase through the action of Rho-kinase. MYPT1 is localized on stress fibers and is distributed close to the cell membrane and at cell-cell contacts to regulate myosin phosphatase activity. It is phosphorylated cooperatively by ROCK1 and CDC42BP on Thr-696 that inhibits PP1C activity by an intramolecular mechanism. Mutations in PPP1R12A are involved in Genitourinary and/or brain malformation syndrome (GUBS), an autosomal dominant syndrome that is characterized by multiple congenital anomalies including urogenital malformations and brain abnormalities. Five isoforms of MYPT1 have been described that are produced by alternative splicing. (Ref.: Qiao, Y-N., et Al. (2014). J. Biol. Chem. 289(32); 22512-22523). |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
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| Presentation | Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mm NaCl with 0.05% sodium azide |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-phospho-MYPT1 (Thr696), Cat. No. ABS45, is a rabbit polyclonal antibody that detects MYPT1 phosphorylated on Threonine 696 and is tested for use in Western Blotting and Peptide Inhibition Assay. |
| Key Applications |
|
| Application Notes | Peptide Inhibition Assay: Target band detection in a lysate from HeLa cells treated with Calyculin (50 nM) and Okadaic Acid (500 nM) was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user. |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated linear peptide corresponding to 11 amino acids surrounding phosphothreonine 696 of human MYPT1. |
| Epitope | Phosphorylated Thr696 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | This rabbit polyclonal antibody specifically detects MYPT1 phosphorylated on threonine 696. |
| Species Reactivity |
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| Species Reactivity Note | Human. Predicted to react with Xenopus, Rat, Canine, Primate, Chicken, Mouse, Horse based on 100% sequence homology. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | Myosin phosphatase target subunit 1, which is also called the myosin-binding subunit of myosin phosphatase, is one of the subunits of myosin phosphatase. Myosin phosphatase regulates the interaction of actin and myosin downstream of the guanosine triphosphatase Rho. The small guanosine triphosphatase Rho is implicated in myosin light chain (MLC) phosphorylation, which results in contraction of smooth muscle and interaction of actin and myosin in nonmuscle cells.The guanosine triphosphate (GTP)-bound, active form of RhoA (GTP.RhoA) specifically interacted with the myosin-binding subunit (MBS) of myosin phosphatase, which regulates the extent of phosphorylation of MLC. Rho-associated kinase (Rho-kinase), which is activated by GTP. RhoA, phosphorylated MBS and consequently inactivated myosin phosphatase. Overexpression of RhoA or activated RhoA in NIH 3T3 cells increased phosphorylation of MBS and MLC. Thus, Rho appears to inhibit myosin phosphatase through the action of Rho-kinase. Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq] |
| Gene Symbol |
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| Modifications |
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| Purification Method | Affinity Purfied |
| UniProt Number |
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| UniProt Summary | FUNCTION: Regulates myosin phosphatase activity. SUBUNIT STRUCTURE: PP1 comprises a catalytic subunit, PPP1CA, PPP1CB or PPP1CC, and one or several targeting or regulatory subunits. PPP1R12A mediates binding to myosin. Interacts with ARHA and CIT By similarity. Binds PPP1R12B, ROCK1 and IL16. SUBCELLULAR LOCATION: Cytoplasm. Note: Along actomyosin filaments and stress fibers. PTM: Phosphorylated by CIT (Rho-associated kinase) By similarity. Phosphorylated cooperatively by ROCK1 and CDC42BP on Thr-696. Phosphorylated on upon DNA damage, probably by ATM or ATR. SEQUENCE SIMILARITIES: Contains 6 ANK repeats. |
| Molecular Weight | ~130 kDa observed; 115.28 kDa calculated. Uncharacterized bands may be observed in some lysate(s). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in HeLa cells treated with Calyculin and Okadaic Acid. Western Blotting Analysis: A 1:500 dilution of this antibody detected phosphorylated MYPT1 (Thr696) in lysate from HeLa cells treated for 30 minutes with Calyculin (50 nM) and Okadaic Acid (500 nM). |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Recommended storage: +2°C to +8°C. |
| Packaging Information | |
|---|---|
| Material Size | 200 µg |