RecombinantRabMAb

Recombinant Anti-Myf5 antibody [EPR4899] (ab125078)

Submit a review Q&A (3)References (7)
Western blot - Anti-Myf5 antibody [EPR4899] (ab125078)
  • Western blot - Anti-Myf5 antibody [EPR4899] (ab125078)
  • Immunoprecipitation - Anti-Myf5 antibody [EPR4899] (ab125078)
  • Western blot - Anti-Myf5 antibody [EPR4899] (ab125078)
  • Anti-Myf5 antibody [EPR4899] (ab125078)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4899] to Myf5
  • Suitable for: WB, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Myf5 antibody [EPR4899]
    See all Myf5 primary antibodies

  • Description

    Rabbit monoclonal [EPR4899] to Myf5

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Pig

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Myf5 aa 200 to the C-terminus. The exact sequence is proprietary.

  • Positive control

    • WB: Human skeletal muscle, rat muscle, mouse muscle and rat heart tissue lysate.

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab125078 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
WB
1/10000 - 1/50000. Detects a band of approximately 37 kDa (predicted molecular weight: 28 kDa).
IP
1/20.
Notes
WB
1/10000 - 1/50000. Detects a band of approximately 37 kDa (predicted molecular weight: 28 kDa).
IP
1/20.
  • Application notes
    Is unsuitable for ICC/IF.

    • Target

    Images

    • Western blot - Anti-Myf5 antibody [EPR4899] (ab125078)
      Western blot - Anti-Myf5 antibody [EPR4899] (ab125078)
      Anti-Myf5 antibody [EPR4899] (ab125078) at 1/50000 dilution (purified) + Human skeletal muscle tissue lysate at 10 µg

      Secondary
      HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 28 kDa
      Observed band size: 39 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST
    • Western blot - Anti-Myf5 antibody [EPR4899] (ab125078)
      Western blot - Anti-Myf5 antibody [EPR4899] (ab125078)
      All lanes : Anti-Myf5 antibody [EPR4899] (ab125078) at 1/50000 dilution (purified)

      Lane 1 : Rat muscle tissue lysate
      Lane 2 : Mouse muscle tissue lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 28 kDa
      Observed band size: 39 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST
    • Immunoprecipitation - Anti-Myf5 antibody [EPR4899] (ab125078)
      Immunoprecipitation - Anti-Myf5 antibody [EPR4899] (ab125078)

      ab125078 (Purified) at 1/20 dilution (20 µg/ml) immunoprecipitating Myf5 in Human skeletal muscle lysate.

      Lane 1 (input): Human skeletal muscle
      Lane 2 (+): 125078 & Human skeletal muscle lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab125078 in human skeletal muscle lysate
      For western blotting, ab125078 at 1/1000 and VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.
      Blocking and diluting buffer: 5% NFDM /TBST.

    • Western blot - Anti-Myf5 antibody [EPR4899] (ab125078)
      Western blot - Anti-Myf5 antibody [EPR4899] (ab125078)
      All lanes : Anti-Myf5 antibody [EPR4899] (ab125078) at 1/10000 dilution (unpurified)

      Lane 1 : Human skeletal muscle tissue lysate
      Lane 2 : Rat muscle tissue lysate
      Lane 3 : Mouse muscle tissue lysate
      Lane 4 : Rat heart tissue lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size: 28 kDa

    • Anti-Myf5 antibody [EPR4899] (ab125078)
      Anti-Myf5 antibody [EPR4899] (ab125078)

    References (7)

    Publishing research using ab125078? Please let us know so that we can cite the reference in this datasheet.

    ab125078 has been referenced in 7 publications.

    • Chaudhary P  et al. High altitude mediated skeletal muscle atrophy: Protective role of curcumin. Biochimie 156:138-147 (2019). PubMed: 30347230
    • Skrzypek K  et al. SNAIL is a key regulator of alveolar rhabdomyosarcoma tumor growth and differentiation through repression of MYF5 and MYOD function. Cell Death Dis 9:643 (2018). PubMed: 29844345
    • Hai T  et al. Pilot study of large-scale production of mutant pigs by ENU mutagenesis. Elife 6:N/A (2017). WB ; Pig . PubMed: 28639938
    • Llewellyn KJ  et al. Myogenic differentiation of VCP disease-induced pluripotent stem cells: A novel platform for drug discovery. PLoS One 12:e0176919 (2017). WB ; Human . PubMed: 28575052
    • Tenente IM  et al. Myogenic regulatory transcription factors regulate growth in rhabdomyosarcoma. Elife 6:N/A (2017). WB, ICC/IF . PubMed: 28080960
    • Koltai E  et al. SIRT1 may play a crucial role in overload-induced hypertrophy of skeletal muscle. J Physiol 595:3361-3376 (2017). PubMed: 28251652
    • Torrado M  et al. Pitx2c is reactivated in the failing myocardium and stimulates myf5 expression in cultured cardiomyocytes. PLoS One 9:e90561 (2014). WB ; Human, Pig . PubMed: 24595098

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Question

    Searching for a Myf5 ab tested in ICC and flow cytometry

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    Answer

    Thank you for contacting us today.

    As discussed, we do not have a Myf5 antibody tested in ICC and flow cytometry. However, you will be able to get this product for free with any primary antibody purchase. Just quote the code RABMAB-XBSMG with your order.

    Please do not hesitate to contact me should you have further questions.

    Read More

    Question

    Hola
    La citometria tengo que hacerla con los dos a la vez, aunque luego por motivos tecnicos y de control las haga por separado, pero necesito que sea una doble tincion.
    Tal vez si se usan dos anti-rabbit de longitudes muy lejanas consigo detectarlos sin problema, incluso si teneis secundarios altamente especificos contra las Igg de cada uno de ellos.
    Que puedo hacer con esto ultimo???
    Muchas gracias, espero tu respuesta.
    Saludos,

    Read More
    Answer

    Gracias por tu respuesta.

    Si estas interesada en usar los dos anticuerpos propuestos para citometría, ten en cuenta que el anti-Myf5 no se ha testado en esta técnica, y por tanto no podemos asegurar que baya a funcionar. Por eso te proponía que lo testarais primero por separado, participando en el sistema Abreviews, y si os da buenos resultados lo probéis entonces para la doble tinción.

    Lo que no aconsejo de ninguna manera es hacer la doble tinción directamente, puesto que en caso de no salir, las causas pueden ser múltiples, y una de ellas que el anti-Myf5 no sea válido para esta técnica.

    Respecto al secundario, podríais usar un Mouse anti –Rabbit conjugado con FITC, y un Goat anti-Rabbit conjugado con APC – Cy7. De esta manera se evita reactividad cruzada, y no solapan en el espectro de emisión.

    Si quieres echar un vistazo a los secundarios propuestos te dejo sus enlaces a las datasheets:

    https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG1-heavy-chain-AllophycocyaninCy7-pre-adsorbed-ab130785.html

    https://www.abcam.com/Mouse-monoclonal-4E3-Secondary-Antibody-to-Human-IgG1-FITC-ab99772.html

    Espero que estas soluciones te sean útiles. Si tienes cualquier otra consulta no dudes en preguntarnos.

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    Question

    Hola,
    Estoy interesado en anticuerpos que reconozcan la proteína pax-7 y myf-5 en musculo de ratón. Quiero usar los dos anticuerpos simultáneamente. ¿Tendríais todos los reactivos? ¿Primarios conjugados? ¿o un par primario/secundario que no de lugar a reactividad cruzada? Si tuvierais algún protocolo lo agradecería.
    Muchas gracias

    Read More
    Answer

    Gracias por contactarnos.



    He llevado a cabo una búsqueda en nuestro catálogo de los anticuerpos solicitados. Contra PAX7 tenemos un anticuerpo, creado en conejo y monoclonal, que se ha testado en ratones por Citometría, no está conjugado, por lo que sería necesario usar un secundario apropiado:

    https://www.abcam.com/PAX7-antibody-EPR3353-ab92317.html

    Para detectar Myf5 en ratón hay una posibilidad, pero lamentablemente no se ha testado en Citometría. También es un monoclonal de conejo. Si os interesara usarlo en Citometría podéis participar en nuestro sistema de evaluación e anticuerpos (Abreview), por el cual si usáis un anticuerpo en una especie / aplicación no testada, os beneficiaríais de un anticuerpo gratuito al compartir con nosotros los resultados en forma de Abreview.

    https://www.abcam.com/Myf5-antibody-EPR4899-ab125078.html

    Para saber más sobre este programa:

    https://www.abcam.com/abreviews

    Yo te recomendaría realizar dos ensayos de Citometría separados, así podrías usar el mismo secundario en ambos casos, y probar el anti-Myf5 en esta técnica.

    Tenemos en la web numerosos protocolos para las distintas aplicaciones en las que pueden usarse nuestros productos, te copio el enlace al protocolo de Citometría por si pudiera resultarte útil:

    https://www.abcam.com/index.html?pageconfig=popular_protocols

    https://www.abcam.com/index.html?pageconfig=resource&rid=11381

    Espero haberte ayudado. En cualquier caso no dudes en contactarme de nuevo si necesitaras más información.

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