Anti-Myosin light chain 3 antibody [MLM527] (ab680)

Overview

  • Product nameAnti-Myosin light chain 3 antibody [MLM527]
    See all Myosin light chain 3 primary antibodies
  • Description
    Mouse monoclonal [MLM527] to Myosin light chain 3
  • SpecificityThis antibody reacts with ventricle myosin light chain I.
  • Tested applicationsSuitable for: IHC-P, ELISA, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Cow, Human, Pig
  • Immunogen

    Human ventricle myosin light chain I.

Properties

Applications

Our Abpromise guarantee covers the use of ab680 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 a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with EDTA pH 9 before commencing with IHC staining protocol.
ELISA Use at an assay dependent concentration.
WB 1/1 - 1/100. Detects a band of approximately 25 kDa (predicted molecular weight: 22 kDa).
ICC/IF Use at an assay dependent concentration.

Target

  • FunctionRegulatory light chain of myosin. Does not bind calcium.
  • Involvement in diseaseDefects in MYL3 are the cause of cardiomyopathy familial hypertrophic type 8 (CMH8) [MIM:608751]. 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. CMH8 inheritance can be autosomal dominant or recessive.
    Defects in MYL3 are the cause of cardiomyopathy familial hypertrophic with mid-left ventricular chamber type 1 (MVC1) [MIM:608751]. MVC1 is a very rare variant of familial hypertrophic cardiomyopathy, characterized by mid-left ventricular chamber thickening.
  • Sequence similaritiesContains 3 EF-hand domains.
  • Post-translational
    modifications
    The N-terminus is blocked.
    N-terminus is methylated by METTL11A/NTM1.
  • Information by UniProt
  • Database links
  • Alternative names
    • Cardiac myosin light chain 1 antibody
    • CMH8 antibody
    • CMLC1 antibody
    • ELC of myosin antibody
    • Essential light chain of myosin antibody
    • MLC1SB antibody
    • MLC1V antibody
    • MYL3 antibody
    • MYL3_HUMAN antibody
    • Myosin light chain 1 antibody
    • Myosin light chain 1 slow twitch muscle B/ventricular isoform antibody
    • Myosin light chain 1 slow, B antibody
    • Myosin light chain 3 antibody
    • Myosin, light chain 1, ventricular antibody
    • myosin, light chain 3, alkali; ventricular, skeletal, slow antibody
    • myosin, light polypeptide 3, alkali antibody
    • myosin, light polypeptide 3, alkali; ventricular, skeletal, slow antibody
    • OTTHUMP00000165922 antibody
    • Slow skeletal ventricular myosin alkali light chain 3 antibody
    • slow-twitch muscle B/ventricular isoform antibody
    • Ventricular/slow twitch myosin alkali light chain antibody
    • VLC1 antibody
    see all

Anti-Myosin light chain 3 antibody [MLM527] images

  • Procedure:
    Coat microplate with porcine cardiac myosin light chain I
    Wash 3 times
    Block with 1% BSA-PBS
    Add MLM527 diluted with BSA-PBS
    Incubate for 1 h at room temp
    Wash 3 times
    Add HRP-labelled anti-mouse IgG diluted with 1% BSA-PBS
    Incubate for 1 h at room temp
    Wash 3 times
    Add TMBZ solution
    Incubate few min
    Measure A450 nm

  • ab680 (1µg/ml) staining myosin light chain in human heart (left ventricle), using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of cardiomyocytes.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • All lanes : Anti-Myosin light chain 3 antibody [MLM527] (ab680) at 1 µg/ml

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Heart (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 22 kDa
    Observed band size : 25 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 50 kDa,63 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds
  • ab680 staining Myosin light chain 3 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 (1/2000 in blocking buffer) for 1 hour. An HRP-conjugated Goat anti-mouse polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

References for Anti-Myosin light chain 3 antibody [MLM527] (ab680)

This product has been referenced in:
  • Singh AP & Archer TK Analysis of the SWI/SNF chromatin-remodeling complex during early heart development and BAF250a repression cardiac gene transcription during P19 cell differentiation. Nucleic Acids Res 42:2958-75 (2014). IHC-P ; Mouse . Read more (PubMed: 24335282) »
  • Peng Y  et al. Critical roles of miRNA-mediated regulation of TGFß signalling during mouse cardiogenesis. Cardiovasc Res 103:258-67 (2014). Mouse . Read more (PubMed: 24835278) »

See all 11 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Username

Abcam user community

Verified customer

Submitted Feb 19 2013

Thank you for your patience. I have received a reply from the originator of ab680. This antibody is raised against the ventricular myosin light chain I as described in the article "Kato H et al. Clin Chem 38, 170-171 1992". The epitope of this ant...

Read More

ab680 cannot be used to probe skeletal isoforms or the cardiac/slow regulatory myosin light chain (VLC2).

The datasheet of this antibody states that it is suitable for ICC. One of our customers has submitted a review, please take a look at her report on the review top panel.

We have not investigated the reactivity of Ab680 with Non-muscle myosin light chain. However, previous studies with other similar antibodies show very low cross-reactivity to other myosin light chains. Therefore, we assume that this antibody does not r...

Read More

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