Overview

  • Product name
    Anti-Myosin Light Chain 2 antibody [EPR3741]
    See all Myosin Light Chain 2 primary antibodies
  • Description
    Rabbit monoclonal [EPR3741] to Myosin Light Chain 2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Myosin Light Chain 2 aa 150-250. The exact sequence is proprietary.

  • Positive control
    • WB: Recombinant Human Myosin Light Chain 2 protein (ab117178), Fetal heart, fetal skeletal muscle, Human heart, mouse heart and rat heart lysates IHC-P: Human heart muscle
  • General notes

     

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab92721 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-Fr Use at an assay dependent concentration. PubMed: 23365102
WB 1/1000 - 1/5000. Predicted molecular weight: 19 kDa.
IP 1/50.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

The use of an HRP/AP polymerized secondary antibody is recommended.

  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Involvement in disease
      Defects in MYL2 are the cause of cardiomyopathy familial hypertrophic type 10 (CMH10) [MIM:608758]. Familial hypertrophic cardiomyopathy is a hereditary heart disorder characterized by ventricular hypertrophy, which is usually asymmetric and often involves the interventricular septum. The symptoms include dyspnea, syncope, collapse, palpitations, and chest pain. They can be readily provoked by exercise. The disorder has inter- and intrafamilial variability ranging from benign to malignant forms with high risk of cardiac failure and sudden cardiac death.
      Defects in MYL2 are the cause of cardiomyopathy familial hypertrophic with mid-left ventricular chamber type 2 (MVC2) [MIM:608758]. MVC2 is a very rare variant of familial hypertrophic cardiomyopathy, characterized by mid-left ventricular chamber thickening.
    • Sequence similarities
      Contains 3 EF-hand domains.
    • Post-translational
      modifications
      N-terminus is methylated by METTL11A/NTM1.
    • Information by UniProt
    • Database links
    • Alternative names
      • Cardiac myosin light chain-2 antibody
      • Cardiac ventricular myosin light chain 2 antibody
      • CMH10 antibody
      • MLC 2v antibody
      • MLC-2 antibody
      • MLC-2v antibody
      • MLC2 antibody
      • MLRV_HUMAN antibody
      • MYL 2 antibody
      • MYL2 antibody
      • Myosin light chain 2 regulatory cardiac slow antibody
      • Myosin light polypeptide 2 regulatory cardiac slow antibody
      • Myosin regulatory light chain 2 antibody
      • Myosin regulatory light chain 2 ventricular/cardiac muscle isoform antibody
      • Regulatory light chain of myosin antibody
      • RLC of myosin antibody
      • Slow cardiac myosin regulatory light chain 2 antibody
      • ventricular/cardiac muscle isoform antibody
      see all

    Images

    • All lanes : Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721) at 1/1000 dilution

      Lane 1 : Human fetal heart tissue lysate
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 3 : 293T (Human embryonic kidney epithelial cell), whole cell lysate
      Lane 4 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 19 kDa
      Observed band size: 19 kDa


      Exposure time: 10 seconds


      Blocking/Diluting buffer and concentration 5% NFDM/TBST

      Myosin light chain-2 (MYL2, also called MLC-2) is an ~19kDa sarcomeric protein that belongs to the EF-hand calcium binding protein superfamily and exists as three major isoforms encoded by three distinct genes in mammalian striated muscle (PMID: 26074085). Although some cell lines are reported to be MYL2 positive, such as HeLa, 293T and MCF7 (PMID: 21487036, 21487036, 23093227), ab92721 only works in muscle and heart tissues according to our internal testing.

    • All lanes : Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721) at 1/10000 dilution

      Lane 1 : Fetal heart lysate
      Lane 2 : Fetal skeletal muscle lysate
      Lane 3 : Human heart lysate
      Lane 4 : Mouse heart lysate
      Lane 5 : Rat heart lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 19 kDa

    • ab92721, at a 1/100 dilution, staining Myosin Light Chain 2 in paraffin embedded Human heart muscle by Immunohistochemistry.
    • All lanes : Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721) at 1/10000 dilution

      Lane 1 : Recombinant Human Myosin Light Chain 2 protein (ab79185) at 0.1 µg
      Lane 2 : Recombinant Human Myosin Light Chain 2 protein (ab79185) at 0.01 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 19 kDa


      Exposure time: 4 minutes
    • ab92721 showing positive staining in Heart muscle tissue.

    • ab92721 showing positive staining in Skeletal muscle tissue.

    • ab92721 showing negative staining in Normal tonsil tissue.

    • ab92721 showing negative staining in Normal brain tissue.

    • ab92721 showing negative staining in Normal liver tissue.

    • ab92721 showing negative staining in Normal kidney tissue.

    • ab92721 showing negative staining in Normal breast tissue.

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:
    • Hodatsu A  et al. Impact of cardiac myosin light chain kinase gene mutation on development of dilated cardiomyopathy. ESC Heart Fail 6:406-415 (2019). Read more (PubMed: 30690923) »
    • Irion CI  et al. Osteopontin Expression in Cardiomyocytes Is Increased in Pediatric Patients With Sepsis or Pneumonia. Front Physiol 9:1779 (2018). Read more (PubMed: 30618794) »
    See all 16 Publications for this product

    Customer reviews and Q&As

    1-10 of 12 Abreviews or Q&A

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% TritonX100 in PBS
    Specification
    HeLa
    Fixative
    Paraformaldehyde

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Oct 27 2015

    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Ileon)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    15 µg
    Specification
    Ileon
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Sep 30 2015

    Western blot

    Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (15)
    Sample
    Rat Tissue lysate - whole (Skeletal muscle)
    Specification
    Skeletal muscle
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted May 22 2013

    Western blot

    Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (15)
    Sample
    Mouse Tissue lysate - whole (Heart)
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted May 22 2013

    Western blot

    Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (15)
    Sample
    Human Tissue lysate - whole (Heart)
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted May 22 2013

    Answer

    Thank you for contacting us with your questions.

    The IHC images on the datasheets for ab92721 and ab137063 were produced in the Epitomics lab, and they weren't taken from publications so there are no references. Ab92721 was used in one publication-

    http://www.ncbi.nlm.nih.gov/pubmed/22037191?dopt=Abstract

    We only have full size vials of our antibodies, so we don't have samples to send out. However, we do have a promotion on rabbit monoclonal antibodies that is valid until December 16. For this promotion, if you purchase any primary antibody you can receive a free rabbit monoclonal. Since both of these antibodies are rabbit monoclonals (also called RabMabs), you could purchase one and then receive the other for free. If you'd like to take advantage of this offer, please include the code RABMAB-XBSMG with your order.

    I hope that this information will be useful, but please let me know if you have further questions or need anything else and I'll be happy to help.

    Read More

    Answer

    Are you interested in an antibody against a different cardiac marker such as GATA4? The antibody ab86371 looks good.

    Click here (or use the following: https://www.abcam.com/index.html?datasheet=86371).

    The antibody ab8295, which you used before successfully, is a cardiac troponin T antibody. Do you want another vial of that, to replace the other troponin T antibody, ab33589? If not, then I suggest ab89594.

    Click here (or use the following: https://www.abcam.com/index.html?datasheet=89594).

    I look forward to your reply.

    Read More

    Answer

    Thank you for bringing this to our attention. If I cannot offer a useful suggestion to improve your results, and if the antibodies are covered by our guarantee, I will send replacements.

    I will first need a few details of your experiment and your order.

    1. Can you please tell me when you ordered the antibodies? If you have a purchase order number or the name of the person who placed the order, that will help me find it in our records.

    2. Please describe your samples: what is the species and cell type or tissue, and how were the lysates prepared? Did you use protease inhibitors?

    3. How much protein did you load per lane of the gel?

    4. Are you confident that the transfer from gel to blot was successful?

    5. What is the blocking solution? (For example, 5% milk in TBS or 1% BSA, etc.).

    6. You mentioned using the antibodies as advised on the datasheets. What dilutions did you try for each antibody?

    7. Are confident the secondary antibody is effective?

    I look forward to your reply.

    Read More

    Answer

    I have issued a free of charge replacement for one vial of ab92721. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

    Read More

    Question
    Answer

    Thank you for contacting Abcam regarding ab92721 and ab89594. I have received the following protocol information from the laboratory: Western Blotting Protocol Solutions and Reagents 2X Laemmli Sample Buffer: 62.5 mM Tris-HCl (pH 6.8), 25% glycerol, 2% SDS, 0.01% Bromophenol Blue, 710 mM beta-mercaptoethanol TBST (50 mM Tris, pH 7.6, 150 mM NaCl, 0.1% Tween-20) TBST is Tris Base Saline buffer with 0.1% Tween-20 5% bovine serum albumin (BSA): Use 25 g in 500 ml TBST Goat anti-rabbit HRP antibody - suggest ab6721 Goat anti-mouse-HRP - suggest ab6789 Chemiluminesence reagents; such as ECL materials and film or imaging system for detection Protocol Run SDS-PAGE using 10-60ug heart lysate and transfer to nitrocellulose membrane. Block nitrocellulose for 1 hour at room temperature or overnight at 4°C using 5% BSA Incubate nitrocellulose with appropriate dilutions of primary antibody in 5% BSA overnight at 4°C or for 2 hours at room temperature Wash nitrocellulose with three 5-min washes using TBST. Incubate nitrocellulose with goat anti-rabbit (or goat anti-mouse) HRP antibody, diluted to 1:500 to 100,000 in 5% BSA for 1-2 hours at room temperature. Wash nitrocellulose in 3 washes of TBST, then rinse in TBS prior to addition of chemiluminesence reagents. Remove excess chemiluminescence reagent and sandwich nitrocellulose blot in any type of transparent plastic wrap (plastic cling wrap, transparent sheet protector, etc.). Acquire image using darkroom development techniques. I hope this information is helpful.  Please to not hesitate to contact us if you have any additional questions.

    Read More

    1-10 of 12 Abreviews or Q&A

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

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