RabMAb

Anti-Myosin Phosphatase 1+Myosin Phosphatase 2抗体[YE336] (ab32519)

概述

  • 产品名称Anti-Myosin Phosphatase 1+Myosin Phosphatase 2抗体[YE336]
  • 描述
    兔单克隆抗体[YE336] to Myosin Phosphatase 1+Myosin Phosphatase 2
  • 特异性This antibody is specific for human Myosin Phosphatase 1 and Myosin Phosphatase 2.
  • 经测试应用适用于: WB, ICC/IF, Flow Cyt, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Myosin Phosphatase 1+Myosin Phosphatase 2 (C terminal).

  • 阳性对照
    • WB: HeLa, Jurkat, 293T, MCF-7, NIH/3T3 and C6 whole cell lysate. ICC/IF: HEK293 and NIH/3T3 cells FC: HEK293 and HeLa cells,
  • 常规说明

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

性能

应用

Our Abpromise guarantee covers the use of ab32519 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/2000. Predicted molecular weight: 110 kDa.

For unpurified use at 1/5000. 

ICC/IF 1/100.
Flow Cyt 1/40.

For unpurified use at 1/100.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/30.

For unpurified use at 1/80. 

靶标

  • 相关性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.
  • 细胞定位Cytoplasmic; along actomyosin filaments and stress fibers.
  • 数据库链接
  • 别名
    • Myosin phosphatase target subunit 1 antibody
    • Myosin phosphatase target subunit 2 antibody
    • Myosin phosphatase targeting subunit 1 antibody
    • Myosin phosphatase targeting subunit 2 antibody
    • MYPT1 antibody
    • MYPT2 antibody
    • PPP1R12A antibody
    • PPP1R12B antibody
    • Protein phosphatase 1 regulatory inhibitor subunit 12A antibody
    • Protein phosphatase 1 regulatory Inhibitor subunit 12B antibody
    • Protein phosphatase 1 regulatory subunit 12A antibody
    • Protein phosphatase 1 regulatory subunit 12B antibody
    • Protein phosphatase myosin binding subunit antibody
    see all

Anti-Myosin Phosphatase 1+Myosin Phosphatase 2 antibody [YE336] 图像

  • All lanes : Anti-Myosin Phosphatase 1+Myosin Phosphatase 2 antibody [YE336] (ab32519) at 1/2000 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : Jurkat whole cell lysate
    Lane 3 : 293T whole cell lysate
    Lane 4 : MCF-7 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 110 kDa
    Observed band size : 110 kDa
    Blocking and Diluting buffer 5% NFDM/TBST
  • Immunocytochemistry/Immunofluorescence analysis of NIH/3T3 cells labelling Myosin Phosphatase 1+Myosin Phosphatase 2 with purified ab32519 at 1/100. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab7291 anti-Tubulin (mouse mAb) followed by ab150120, AlexaFluor®594 goat anti-mouse secondary both at 1/1000. Nuclei were counterstained with DAPI (blue).

    For negative control 1, rabbit primary antibody was used followed by anti-mouse secondary antibody (ab150120). For negative control 2, ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (ab150077).

  • All lanes : Anti-Myosin Phosphatase 1+Myosin Phosphatase 2 antibody [YE336] (ab32519) at 1/2000 dilution

    Lane 1 : NIH/3T3 whole cell lysate
    Lane 2 : C6 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 110 kDa
    Observed band size : 110 kDa
    Blocking and Diluting buffer 5% NFDM/TBST
  • Anti-Myosin Phosphatase 1+Myosin Phosphatase 2 antibody [YE336] (ab32519) at 1/5000 dilution (unpurified) + 293T cell lysate.

    Predicted band size : 110 kDa
    Observed band size : 110 kDa
  • Flow cytometry analysis of HeLa cells labelling Myosin Phosphatase 1+Myosin Phosphatase 2 (red) with purified ab32519 at dilution of 1/40. The secondary antibody used was goat anti rabbit IgG (FITC) at 1/500. Cells were fixed with 4% paraformaldehyde. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • ab32519 at 1/30 dilution immunoprecipitating Myosin Phosphatase 1+Myosin Phosphatase 2 in HeLa whole cell lysate observed at 110 KDa (lanes 1 and 2).

    Lane 1 (input): HeLa whole cell lysate 10ug

    Lane 2 (+): ab32519 + HeLa whole cell lysate

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32519 in HeLa whole cell lysate

    For western blotting, ab32519 was used followed by VeriBlot for IP secondary antibody (HRP) (ab131366) as the secondary antibody at a dilution of 1/10,000.

    Blocking and Diluting buffer and concentration: 5% NFDM/TBST.

  • ab32519 staining mouse fibroblast cells by ICC/IF. Cells were PFA fixed, permeabilized in Triton X-100 and blocked in 1% BSA for 30 minutes at 25°C. The primary antibody was diluted 1/200 and incubated with sample for 4 hours at 25°C. An Alexa Fluor® 488 conjugated goat monoclonal to rabbit, diluted 1/500 was used as the secondary.

    See Abreview

  • Overlay histogram showing HEK293 cells stained with unpurified ab32519 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32519, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Anti-Myosin Phosphatase 1+Myosin Phosphatase 2 antibody [YE336] (ab32519)参考文献

This product has been referenced in:
  • Yin X  et al. Proteomics analysis of the cardiac myofilament subproteome reveals dynamic alterations in phosphatase subunit distribution. Mol Cell Proteomics 9:497-509 (2010). WB, IHC-Fr ; Mouse, Rat . Read more (PubMed: 20037178) »
  • Kogata N  et al. Integrin-linked kinase controls vascular wall formation by negatively regulating Rho/ROCK-mediated vascular smooth muscle cell contraction. Genes Dev 23:2278-83 (2009). WB ; Mouse . Read more (PubMed: 19797768) »

See all 4 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (7.5%)
Sample Rabbit Cell lysate - whole cell (NSCLC)
Specification NSCLC
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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提交于 Sep 27 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (mouse fibroblast)
Specification mouse fibroblast
Fixative Paraformaldehyde
Permeabilization Yes - TritonX-100
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Abcam user community

Verified customer

提交于 Oct 08 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"