Phospho-MYPT1 (Thr853) Antibody - #AF5445

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Product: Phospho-MYPT1 (Thr853) Antibody
Catalog: AF5445
Description: Rabbit polyclonal antibody to Phospho-MYPT1 (Thr853)
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Zebrafish, Horse, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 115~140 kDa; 115kD(Calculated).
Uniprot: O14974
RRID: AB_2837929

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 100ul $280 In stock
 200ul $350 In stock

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MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Zebrafish(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(88%)
Clonality:
Polyclonal
Specificity:
Phospho-MYPT1 (Thr853) Antibody detects endogenous levels of MYPT1 only when phosphorylated at Thr853.
RRID:
AB_2837929
Cite Format: Affinity Biosciences Cat# AF5445, RRID:AB_2837929.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

M130; MBS; Myosin binding subunit; Myosin phosphatase target subunit 1; Myosin phosphatase targeting subunit 1; Myosin phosphatase-targeting subunit 1; MYPT1; MYPT1_HUMAN; PPP1R12A; Protein phosphatase 1 regulatory inhibitor subunit 12A; Protein phosphatase 1 regulatory subunit 12A; Protein phosphatase 1, regulatory (inhibitor) subunit 12A; Protein phosphatase myosin binding subunit; Protein phosphatase myosin-binding subunit;

Immunogens

Immunogen:
Uniprot:

O14974(MYPT1_HUMAN) >>Visit HPA database.

Gene(ID):
PPP1R12A(4659)
Expression:
O14974 MYPT1_HUMAN:

Expressed in striated muscles, specifically in type 2a fibers (at protein level).

Description:
Heterotetramerization is mediated by the interaction between a coiled-coil of PRKG1 and the leucine/isoleucine zipper of PPP1R12A/MBS, the myosin-binding subunit of the myosin phosphatase.
Sequence:
MKMADAKQKRNEQLKRWIGSETDLEPPVVKRQKTKVKFDDGAVFLAACSSGDTDEVLKLLHRGADINYANVDGLTALHQACIDDNVDMVKFLVENGANINQPDNEGWIPLHAAASCGYLDIAEFLIGQGAHVGAVNSEGDTPLDIAEEEAMEELLQNEVNRQGVDIEAARKEEERIMLRDARQWLNSGHINDVRHAKSGGTALHVAAAKGYTEVLKLLIQAGYDVNIKDYDGWTPLHAAAHWGKEEACRILVDNLCDMEMVNKVGQTAFDVADEDILGYLEELQKKQNLLHSEKRDKKSPLIESTANMDNNQSQKTFKNKETLIIEPEKNASRIESLEQEKVDEEEEGKKDESSCSSEEDEEDDSESEAETDKTKPLASVTNANTSSTQAAPVAVTTPTVSSGQATPTSPIKKFPTTATKISPKEEERKDESPATWRLGLRKTGSYGALAEITASKEGQKEKDTAGVTRSASSPRLSSSLDNKEKEKDSKGTRLAYVAPTIPRRLASTSDIEEKENRDSSSLRTSSSYTRRKWEDDLKKNSSVNEGSTYHKSCSFGRRQDDLISSSVPSTTSTPTVTSAAGLQKSLLSSTSTTTKITTGSSSAGTQSSTSNRLWAEDSTEKEKDSVPTAVTIPVAPTVVNAAASTTTLTTTTAGTVSSTTEVRERRRSYLTPVRDEESESQRKARSRQARQSRRSTQGVTLTDLQEAEKTIGRSRSTRTREQENEEKEKEEKEKQDKEKQEEKKESETSREDEYKQKYSRTYDETYQRYRPVSTSSSTTPSSSLSTMSSSLYASSQLNRPNSLVGITSAYSRGITKENEREGEKREEEKEGEDKSQPKSIRERRRPREKRRSTGVSFWTQDSDENEQEQQSDTEEGSNKKETQTDSISRYETSSTSAGDRYDSLLGRSGSYSYLEERKPYSSRLEKDDSTDFKKLYEQILAENEKLKAQLHDTNMELTDLKLQLEKATQRQERFADRSLLEMEKRERRALERRISEMEEELKMLPDLKADNQRLKDENGALIRVISKLSK

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Horse
100
Dog
100
Zebrafish
100
Chicken
100
Rabbit
100
Xenopus
88
Pig
0
Bovine
0
Sheep
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O14974 As Substrate

Site PTM Type Enzyme
K9 Acetylation
S20 Phosphorylation
T22 Phosphorylation
T34 Phosphorylation
Y68 Phosphorylation
T234 Phosphorylation
K244 Ubiquitination
K285 Ubiquitination
K286 Ubiquitination
S292 Phosphorylation
S299 Phosphorylation
S304 Phosphorylation
S313 Phosphorylation
S336 Phosphorylation
S365 Phosphorylation
T406 Phosphorylation
T408 Phosphorylation
S409 Phosphorylation
T416 Phosphorylation
T417 Phosphorylation
T419 Phosphorylation
S422 Phosphorylation
S432 Phosphorylation P06493 (CDK1)
K442 Acetylation
T443 Phosphorylation O75116 (ROCK2)
S445 Phosphorylation O60285 (NUAK1) , O95835 (LATS1)
Y446 Phosphorylation
T453 Phosphorylation
K456 Acetylation
K460 Acetylation
T464 Phosphorylation
S470 Phosphorylation
S472 Phosphorylation O60285 (NUAK1)
S473 Phosphorylation P06493 (CDK1)
S477 Phosphorylation
S478 Phosphorylation
S479 Phosphorylation
S489 Phosphorylation
Y496 Phosphorylation
T500 Phosphorylation
S507 Phosphorylation
T508 Phosphorylation
S509 Phosphorylation
S520 Phosphorylation
S525 Phosphorylation
T529 Phosphorylation
K539 Methylation
S541 Phosphorylation
S542 Phosphorylation
S547 Phosphorylation
T548 Phosphorylation
Y549 Phosphorylation
K551 Methylation
S552 Phosphorylation
S554 Phosphorylation
T571 Phosphorylation
T575 Phosphorylation
S578 Phosphorylation
S585 Phosphorylation
S601 Phosphorylation P06493 (CDK1)
S607 Phosphorylation
S608 Phosphorylation
T609 Phosphorylation
S618 Phosphorylation
T631 Phosphorylation
T645 Phosphorylation
T647 Phosphorylation
T652 Phosphorylation
S668 Phosphorylation
Y669 Phosphorylation
T671 Phosphorylation
S678 Phosphorylation
S680 Phosphorylation
S692 Phosphorylation Q13976 (PRKG1)
S695 Phosphorylation P17612 (PRKACA) , Q13976 (PRKG1)
T696 Phosphorylation Q09013 (DMPK) , P04049 (RAF1) , Q13464 (ROCK1) , P17612 (PRKACA) , O75116 (ROCK2) , Q13418 (ILK)
T700 Phosphorylation
T702 Phosphorylation
T710 Phosphorylation Q13418 (ILK)
S716 Phosphorylation
T717 Phosphorylation
T719 Phosphorylation
K743 Acetylation
T748 Phosphorylation
S749 Phosphorylation
Y754 Phosphorylation
K755 Acetylation
Y758 Phosphorylation
S759 Phosphorylation
T761 Phosphorylation
Y762 Phosphorylation
T765 Phosphorylation
Y766 Phosphorylation
Y769 Phosphorylation
S781 Phosphorylation
S790 Phosphorylation
Y792 Phosphorylation
S794 Phosphorylation
S795 Phosphorylation
S802 Phosphorylation
Y810 Phosphorylation
S811 Phosphorylation
S839 Phosphorylation
S852 Phosphorylation P17612 (PRKACA) , O75116 (ROCK2) , Q13976 (PRKG1) , Q9Y5S2 (CDC42BPB) , Q13464 (ROCK1)
T853 Phosphorylation P17612 (PRKACA) , Q13464 (ROCK1) , Q04759 (PRKCQ)
S856 Phosphorylation
T859 Phosphorylation
S862 Phosphorylation
S871 Phosphorylation
T873 Phosphorylation
S877 Phosphorylation
T884 Phosphorylation
S886 Phosphorylation
S888 Phosphorylation
Y890 Phosphorylation
T892 Phosphorylation
S893 Phosphorylation
S894 Phosphorylation
S896 Phosphorylation
Y901 Phosphorylation
S903 Phosphorylation
S908 Phosphorylation
S910 Phosphorylation O60285 (NUAK1)
Y911 Phosphorylation
S912 Phosphorylation
Y913 Phosphorylation
Y920 Phosphorylation
S921 Phosphorylation
S922 Phosphorylation
Y936 Phosphorylation
K945 Ubiquitination
S978 Phosphorylation
S995 Phosphorylation
K1002 Methylation
K1008 Methylation
S1029 Phosphorylation

Research Backgrounds

Function:

Key regulator of protein phosphatase 1C (PPP1C). Mediates binding to myosin. As part of the PPP1C complex, involved in dephosphorylation of PLK1. Capable of inhibiting HIF1AN-dependent suppression of HIF1A activity.

PTMs:

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. In vitro, phosphorylation of Ser-695 by PKA and PKG appears to prevent phosphorylation of the inhibitory site Thr-696, probably mediated by PRKG1. Phosphorylation at Ser-445, Ser-472 and Ser-910 by NUAK1 promotes interaction with 14-3-3, leading to inhibit interaction with myosin light chain MLC2, preventing dephosphorylation of MLC2. May be phosphorylated at Thr-696 by DMPK; may inhibit the myosin phosphatase activity. Phosphorylated at Ser-473 by CDK1 during mitosis, creating docking sites for the POLO box domains of PLK1. Subsequently, PLK1 binds and phosphorylates PPP1R12A.

Subcellular Location:

Cytoplasm. Cytoplasm>Cytoskeleton>Stress fiber.
Note: Also along actomyosin filaments.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in striated muscles, specifically in type 2a fibers (at protein level).

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. Interacts directly with PRKG1. Non-covalent dimer of 2 dimers; PRKG1-PRKG1 and PPP1R12A-PPP1R12A. Interacts with SMTNL1 (By similarity). Interacts with PPP1CB; the interaction is direct. Interacts (when phosphorylated at Ser-445, Ser-472 and Ser-910) with 14-3-3. Interacts with ROCK1 and ROCK2. Interacts with isoform 1 and isoform 2 of ZIPK/DAPK3. Interacts with RAF1. Interacts with HIF1AN. Interacts with NCKAP1L.

Family&Domains:

Heterotetramerization is mediated by the interaction between a coiled-coil of PRKG1 and the leucine/isoleucine zipper of PPP1R12A/MBS, the myosin-binding subunit of the myosin phosphatase.

The KVKF motif mediates interaction with PPP1CB.

Research Fields

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cell motility > Regulation of actin cytoskeleton.   (View pathway)

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cAMP signaling pathway.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Organismal Systems > Circulatory system > Vascular smooth muscle contraction.   (View pathway)

· Organismal Systems > Immune system > Platelet activation.   (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

References

1). Icariin promotes proliferation and osteogenic differentiation of rat adipose-derived stem cells by activating the RhoA-TAZ signaling pathway. Biomedicine & Pharmacotherapy, 2017 (PubMed: 28122303) [IF=7.5]

Application: WB    Species: rat    Sample:

Fig. 3. Icariin treatment increased active RhoA in rASCs. (A) The active and total RhoA protein levels were determined by Western blot analysis on days 0, 1, 3, 5, 7 and 9. The densitometric analysis of active RhoA protein expression was normalized to total RhoA. (B) The p-MYPT1 and GAPDH protein levels were determined by Western blot analysis on days 0, 1, 3, 5, 7 and 9. The densitometric analysis of p-MYPT1 protein expression was normalized to GAPDH. Data are shown as mean  SD (n = 3) in three independent experiments. *P < 0.05 compared to 0 day group.
2). Flavonoids of Coreopsis tinctoria Nutt alleviate the oxidative stress and inflammation of glomerular mesangial cells in diabetic nephropathy via RhoA/ROCK signaling. Journal of Functional Foods, 2022 [IF=5.6]
3). Silencing of ANGPTL8 Alleviates Insulin Resistance in Trophoblast Cells. Frontiers in Endocrinology, 2021 (PubMed: 34163433) [IF=5.2]

Application: WB    Species: human    Sample: HTR-8/SVneo cell

FIGURE 3 | ANGPTL8 activated JNK and ROCK signaling.(C, D) The levels of ROCK I, ROCK II, p-MYPT1 and MYPT1 in HTR-8/SVneo cells.

Application: WB    Species: Human    Sample: HTR-8/SVneo cells

Figure 3 ANGPTL8 activated JNK and ROCK signaling. (A, B) The levels of p-JNK and JNK in HTR-8/SVneo cells with IR, overexpression or silence of ANGPTL8 or/and insulin stimulation. (C, D) The levels of ROCK I, ROCK II, p-MYPT1 and MYPT1 in HTR-8/SVneo cells. (E) Glucose consumption was determined in HTR-8/SVneo cells with ANGPTL8 knockdown, JNK agonist Anisomycin (1 μg/ml) or JNK antagonist SP600125 (20 μM), and insulin stimulation. (F) The levels of insulin signaling related molecules were determined in HTR-8/SVneo cells. (*p < 0.05, **p < 0.01, ns. no significance).
4). Acetylated α-Tubulin Regulated by N-Acetyl-Seryl-Aspartyl-Lysyl-Proline(Ac-SDKP) Exerts the Anti-fibrotic Effect in Rat Lung Fibrosis Induced by Silica. Scientific Reports, 2016 (PubMed: 27577858) [IF=4.6]

Application: WB    Species: rat    Sample:

Figure 4. Effect of Ac-SDKP on col I, α-SMA and α-Ac-Tub in fibroblasts induced by Ang II. (a) The coexpression of α-SMA and α-Ac-Tub in fibroblasts induced by Ang II measured by immunofluorescence. Scale bar=50μm. (b) Effect of Ac-SDKP, valsartan (AT1 inhibitor), TCS HDAC6 20b (specific HDAC6 inhibitor), and Y-27632 (ROCK inhibitor) on fibroblasts measured by Western blot. Data presented as mean±SEM; N=4 independent experiments.
5). Rho GDP dissociation inhibitor α silencing attenuates silicosis by inhibiting RhoA/Rho kinase signalling. EXPERIMENTAL CELL RESEARCH, 2019 (PubMed: 31029634) [IF=3.7]

Application: WB    Species: human    Sample: MRC-5 cells

Fig. 4.| Depletion of RhoGDIα attenuated myofibroblast differentiation by suppression of RhoA/ROCK signalling. (A), Levels of RhoA and phospho-MYPT in MRC-5 cells with RhoGDIα knockdown were determined by western blotting (n = 3).

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