Overview

  • Product name

    Anti-Myosin Phosphatase (phospho T696) antibody
    See all Myosin Phosphatase primary antibodies
  • Description

    Rabbit polyclonal to Myosin Phosphatase (phospho T696)
  • Host species

    Rabbit
  • Specificity

    Detects endogenous levels of Myosin Phosphatase only when phosphorylated at threoine 696.
  • Tested applications

    Suitable for: IHC-P, ELISAmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic phosphopeptide (Human) from around the phosphorylation site of threonine 696 (RSTPQG)

  • Positive control

    • Human breast carcinoma tissue

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride

    Without Mg+2 and Ca+2
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Affinity purified from rabbit antiserum by affinity chromatography using epitope specific phosphopeptide. The antibody against non phosphopeptide was removed by chromatography using non phosphopeptide corresponding to the phosphorylation site.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab59202 in the following tested applications.

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

Application Abreviews Notes
IHC-P Use at an assay dependent concentration.
ELISA 1/5000.

Target

  • Function

    Regulates myosin phosphatase activity.
  • Tissue specificity

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

    Contains 6 ANK repeats.
  • Developmental stage

    Induced by 2-fold during pregnancy, including in abdominus rectus muscle.
  • Domain

    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.
  • Post-translational
    modifications

    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.
  • Cellular localization

    Cytoplasm. Along actomyosin filaments and stress fibers.
  • Information by UniProt
  • Database links

  • Alternative names

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

Images

  • Immunohistochemical analysis of paraffin embedded human breast carcinoma tissue using ab59202 at 1/50 dilution. Samples were untreated (left) or treated with immunizing peptide (right).

References

This product has been referenced in:

  • Yin Y  et al. Expression of MYPT1, CPI-17 and MLC20 in ileum of neonatal mouse NEC model and its significance. Exp Ther Med 14:2221-2227 (2017). Read more (PubMed: 28962146) »
  • Sastre E  et al. Decompensated liver cirrhosis and neural regulation of mesenteric vascular tone in rats: role of sympathetic, nitrergic and sensory innervations. Sci Rep 6:31076 (2016). WB ; Rat . Read more (PubMed: 27484028) »
See all 2 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for your inquiry.

We don't have a polymer kit besides our mouse on mouse polymer kit, ab127055.

https://www.abcam.com/Mouse-on-Mouse-MOM-Polymer-IHC-Kit-ab127055.html

We have ABC kits and EXPOSE kits.

https://www.abcam.com/Mouse-and-Rabbit-Specific-HRPDAB-ABC-detection-IHC-kit-ab64264.html

https://www.abcam.com/index.html?pageconfig=resource&rid=12865

I hope this information helps. Please contact us with any other questions.

Read More

Answer

Thank you for your reply.


This is just to go along with the polymer kit that was used. If you are using polymer detection, simply follow the manufacturers instructions. You can omit this step if not using this type of amplification.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

Read More

Question
Answer

Thank you for contacting Abcam regarding ab59202.



The IHC protocol is as follows:



Deparaffinization

1. Incubate slide at 60℃ for 60 minutes.

2. Deparaffinize in Xylene for 10 minutes and repeat one more times.

3. Hydrate in 100% alcohol for 5 minutes, in 95% alcohol for 5 minutes, in 85% alcohol for 5 minutes, in 75% alcohol for 5 minutes.

4. Dip into Distill Water for 5 minutes.

5. Dip into TBS (50 mM Tris, 100 mM NaCl, pH 7.6), leave for 5 minutes, and repeat two times.



Antigen Retrieval

6. Bring 500 - 2000 ml 10 mM citrate buffer (pH6.0) to the boil in a stainless steel pressure cooker.

7. Put the slide into staining rack and lower into pressure cooker ensuring the slide is well immersed in citrate buffer.

8. When the pressure indicator valve has risen after 3-4 minutes, incubate for 1 minute.

9. Cool the slide naturally to room temperature.

10. Dip into distilled water, leave for 5 minutes, and repeat two times.

11. Dip the slide in TBS for 5 minutes and repeat two times.

12. Immerse slides in 3% H2O2 (in fresh methanol) for 15 minutes at room temperature.

13. Wash with distilled water two times, 5 minutes each time.

14. Wash with TBS (pH 7.6) two times, 5 minutes each time.



Staining with Primary Antibody

15. Dilute primary antibody with 3% BSA in TBS to 1:50. Cover the tissue section on the slide with diluted primary antibody (use 50 – 150μl for each slide).

16. Incubate at 37℃ for 30 minutes or at room temperature for 60 minutes (The optimal incubation time, incubation temperature, and antibody dilution should be determined by the individual laboratory).

17. Wash with TBS two times, 5 minutes each time.



Staining with Secondary Antibody

18. Incubate with 100-200μl Polymer Enhancer. Incubate 30 minutes at 37℃.

19. Wash with TBS for 3 times, 5 minutes each time.

20. Incubate with 100-200μl Polymerized HRP and incubate 30 minutes at 37℃

21. Wash with TBS for 3 times, 5 minutes each time.

22. Add DAB solution and incubate 3-10 minutes (The reaction progress and the optimal time should be determine according to microscope).

23. Wash with distilled water for 2 times, 5 minutes each time.

24. Counterstain sections in hematoxylin if required,wash with distilled water.Immerse slides in 0.1% HCl ethanol for 1-10 seconds, wash with distilled water.

25. Dehydrate through 95% ethanol for 1 minute, 100%



I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela)
Loading amount
120 µg
Specification
Hela
Treatment
10ng/ml IL6 for 30 minutes
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

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Verified customer

Submitted Sep 02 2008

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