Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-MYL9 antibody [EPR13012(2)] (ab191393)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13012(2)] to MYL9
  • Suitable for: WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-MYL9 antibody [EPR13012(2)]
    See all MYL9 primary antibodies

  • Description

    Rabbit monoclonal [EPR13012(2)] to MYL9

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human MYL9 aa 50-150. The exact sequence is proprietary.
    Database link: P24844

  • Positive control

    • WB: HeLa, Hu colon, C6, PC12 and NIH 3T3 cell lysates; Human stomach, colon cancer and uterus lysates; Rat stomach lysate. IHC-P: Human and rat colon tissues. IP: Human stomach 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.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR13012(2)

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab191393 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 1/1000 - 1/2000. Detects a band of approximately 20 kDa (predicted molecular weight: 20 kDa).
IHC-P 1/350 - 1/700. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/40.

Target

  • Function

    Myosin regulatory subunit that plays an important role in regulation of both smooth muscle and nonmuscle cell contractile activity via its phosphorylation. Implicated in cytokinesis, receptor capping, and cell locomotion.

  • Tissue specificity

    Smooth muscle tissues and in some, but not all, nonmuscle cells.

  • Sequence similarities

    Contains 3 EF-hand domains.

  • Post-translational
    modifications

    Phosphorylation increases the actin-activated myosin ATPase activity and thereby regulates the contractile activity. It is required to generate the driving force in the migration of the cells but not necessary for localization of myosin-2 at the leading edge.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 20 kDa myosin light chain antibody
    • Human 20kDa myosin light chain (MLC2) mRNA complete cds antibody
    • LC20 antibody
    • MGC3505 antibody
    • MLC 2 antibody
    • MLC-2C antibody
    • MLC2 antibody
    • MLY 9 antibody
    • MRLC1 antibody
    • MYL9 antibody
    • MYL9_HUMAN antibody
    • Myosin light chain 9 regulatory antibody
    • Myosin light polypeptide 9 regulatory antibody
    • myosin regulatory light chain 1 antibody
    • Myosin regulatory light chain 2 antibody
    • Myosin regulatory light chain 2 smooth muscle isoform antibody
    • Myosin regulatory light chain 9 antibody
    • Myosin regulatory light chain MRLC1 antibody
    • Myosin regulatory light polypeptide 9 antibody
    • Myosin RLC antibody
    • Myosin vascular smooth muscle light chain 2 antibody
    • MYRL2 antibody
    • OTTHUMP00000030857 antibody
    • smooth muscle isoform antibody
    see all

Images

  • Western blot - Anti-MYL9 antibody [EPR13012(2)] (ab191393)
    Western blot - Anti-MYL9 antibody [EPR13012(2)] (ab191393)
    All lanes : Anti-MYL9 antibody [EPR13012(2)] (ab191393) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : MYL9 knockout HeLa cell lysate
    Lane 3 : Human colon cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 20 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab191393 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab191393 Anti-MYL9 antibody [EPR13012(2)] was shown to specifically react with MYL9 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266036 (knockout cell lysate ab256999) was used. Wild-type and MYL9 knockout samples were subjected to SDS-PAGE. ab191393 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MYL9 [EPR13012(2)] antibody (ab191393)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MYL9 [EPR13012(2)] antibody (ab191393)

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling MYL9 with ab191393 at 1/700 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin. Inset: Negative control: using PBS instead of primary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-MYL9 [EPR13012(2)] antibody (ab191393)
    Western blot - Anti-MYL9 [EPR13012(2)] antibody (ab191393)
    All lanes : Anti-MYL9 antibody [EPR13012(2)] (ab191393) at 1/1000 dilution

    Lane 1 : Human stomach lysate
    Lane 2 : Human colon cancer lysate
    Lane 3 : Human uterus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 20 kDa

  • Western blot - Anti-MYL9 [EPR13012(2)] antibody (ab191393)
    Western blot - Anti-MYL9 [EPR13012(2)] antibody (ab191393)
    All lanes : Anti-MYL9 antibody [EPR13012(2)] (ab191393) at 1/1000 dilution

    Lane 1 : C6 cell lysate
    Lane 2 : PC12 cell lysate
    Lane 3 : NIH 3T3 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 20 kDa

  • Western blot - Anti-MYL9 [EPR13012(2)] antibody (ab191393)
    Western blot - Anti-MYL9 [EPR13012(2)] antibody (ab191393)
    Anti-MYL9 antibody [EPR13012(2)] (ab191393) at 1/5000 dilution + rat stomach lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 20 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MYL9 [EPR13012(2)] antibody (ab191393)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MYL9 [EPR13012(2)] antibody (ab191393)

    Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling MYL9 with ab191393 at 1/700 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin. Inset: Negative control: using PBS instead of primary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-MYL9 [EPR13012(2)] antibody (ab191393)
    Immunoprecipitation - Anti-MYL9 [EPR13012(2)] antibody (ab191393)

    Western blot analysis of immunoprecipitation pellet from Human stomach lysate immunoprecipitated using ab191393 at 1/40 dilution (lane 1) or PBS control (lane 2).
    Secondary: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.

References (1)

Publishing research using ab191393? Please let us know so that we can cite the reference in this datasheet.

ab191393 has been referenced in 1 publication.

  • Wang JH  et al. Expression and prognostic significance of MYL9 in esophageal squamous cell carcinoma. PLoS One 12:e0175280 (2017). IHC-P ; Human . PubMed: 28388691

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