Overview

  • Product name

    Anti-Myosin Light Chain 2 antibody [AT3B2]
    See all Myosin Light Chain 2 primary antibodies
  • Description

    Mouse monoclonal [AT3B2] to Myosin Light Chain 2
  • Host species

    Mouse
  • Tested applications

    Suitable for: Flow Cyt, WB, Indirect ELISA, IHC-Pmore details
  • Species reactivity

    Reacts with: Rat, Rabbit, Human
  • Immunogen

    Recombinant full length protein corresponding to Human Myosin Light Chain 2 aa 1-166.
    Database link: P10916

  • Positive control

    • Rat heart extracts.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 11th April 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab89594 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
Flow Cyt Use at an assay dependent concentration.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

WB Use at an assay dependent concentration. Detects a band of approximately 19 kDa (predicted molecular weight: 19 kDa).
Indirect ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

  • Overlay histogram showing A431 cells stained with ab89594 (red line). The cells were fixed with 80% methanol (5 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 (ab89594, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This image was generated using the ascites version of the product.

  • Anti-Myosin Light Chain 2 antibody [AT3B2] (ab89594) at 1/1000 dilution + rat heart extracts at 60 µg

    Secondary
    goat anti-mouse HRP

    Developed using the ECL technique.

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



    This image was generated using the ascites version of the product.

  • ab98594 staining Myosin Light Chain 2 in Rabbit heart tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 60 minutes; antigen retrieval was by heat mediation in a Tris/EDTA buffer. Samples were incubated with primary antibody (1/500 in PBS-T) for 1 hour at 37°C. A HRP-conjugated Goat anti-mouse polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • Anti-Myosin Light Chain 2 antibody [AT3B2] (ab89594) at 1/2000 dilution + Rabbit heart tissue lysate at 10 µg

    Secondary
    HRP conjugated goat anti-mouse at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 19 kDa


    Exposure time: 1 minute


    This image was generated using the ascites version of the product.

    See Abreview

References

This product has been referenced in:

  • Mourino-Alvarez L  et al. A comprehensive study of calcific aortic stenosis: from rabbit to human samples. Dis Model Mech 11:N/A (2018). Read more (PubMed: 29752279) »
  • Russo V  et al. Porous, Ventricular Extracellular Matrix-Derived Foams as a Platform for Cardiac Cell Culture. Biores Open Access 4:374-88 (2015). Read more (PubMed: 26487982) »
See all 3 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
Sample
Rabbit Tissue lysate - whole (Heart)
Loading amount
10 µg
Specification
Heart
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7.5%

Abcam user community

Verified customer

Submitted Feb 22 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Heart)
Specification
Heart
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%

Abcam user community

Verified customer

Submitted Feb 21 2013

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
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (hESC-derived cardiomyocyte)
Specification
hESC-derived cardiomyocyte
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 03 2010

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