Overview

  • Product name
    Anti-Sarcomeric Alpha Actinin antibody [EA-53]
    See all Sarcomeric Alpha Actinin primary antibodies
  • Description
    Mouse monoclonal [EA-53] to Sarcomeric Alpha Actinin
  • Host species
    Mouse
  • Specificity
    According to the following publication, this antibody clone detects the Alpha Actinin 2. However, we have not confirmed this with internal testing if it detects Actinin 1 or 2. https://academic.oup.com/hmg/article/14/12/1587/640357/Alpha-actinin-associates-with-polycystin-2-and
  • Tested applications
    Suitable for: IHC-P, IHC-Fr, ICC/IF, IHC-FoFr, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Goat, Chicken, Hamster, Cow, Cat, Dog, Human, Pig, Xenopus laevis, Fish, Lizard, Snake, Xenopus tropicalis
  • Immunogen

    Full length native protein (purified) corresponding to Rabbit Sarcomeric Alpha Actinin. Purified rabbit skeletal α-actinin.

  • Positive control
    • WB: Mouse and rat skeletal muscle and rat heart tissue lysates. IHC-P: Human skeletal muscle tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab9465 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 1/25 - 1/50. ABC method. We suggest an incubation period of 60 minutes at room temperature. However, depending upon the fixation conditions and the staining system employed, optimal incubation and dilutions should be determined by the user.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 17383364
IHC-FoFr Use at an assay dependent concentration. PubMed: 20019242
WB Use a concentration of 1 µg/ml. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).

Target

  • Function
    F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein.
  • Tissue specificity
    Expressed in both skeletal and cardiac muscle.
  • Involvement in disease
    Defects in ACTN2 are the cause of cardiomyopathy dilated type 1AA (CMD1AA) [MIM:612158]. Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
  • Sequence similarities
    Belongs to the alpha-actinin family.
    Contains 1 actin-binding domain.
    Contains 2 CH (calponin-homology) domains.
    Contains 2 EF-hand domains.
    Contains 4 spectrin repeats.
  • Cellular localization
    Cytoplasm > myofibril > sarcomere > Z line. Colocalizes with MYOZ1 and FLNC at the Z-lines of skeletal muscle.
  • Information by UniProt
  • Database links
  • Alternative names
    • Actin binding protein antibody
    • Actinin alpha 2 antibody
    • ACTN 2 antibody
    • ACTN2 antibody
    • ACTN2_HUMAN antibody
    • Alpha actinin 2 antibody
    • Alpha actinin skeletal muscle antibody
    • Alpha actinin skeletal muscle isoform 2 antibody
    • Alpha-actinin skeletal muscle isoform 2 antibody
    • Alpha-actinin-2 antibody
    • CMD1AA antibody
    • F actin cross linking protein antibody
    • F-actin cross-linking protein antibody
    see all

Images

  • ab9465 staining Sarcomeric Alpha Actinin in mouse heart tissue by Immunohistochemistry (Frozen sections). Serial longitudinal (12 µm) sections from wt hearts were incubated with ab9465. Nuclei were visualized with DAPI.
  • Immunocytochemistry/Immunofluorescence analysis of H9 hESC and CBiPSC6.2 cells labelling Sarcomeric Alpha Actinin (green) with ab9465 at a dilution of 1/200. An Alexa Fluor® 488-conjugated goat anti-mouse IgG was used as the secondary antibody (1/200).

  • Immunofluorescence staining of SNAIL1 and cardiac cell type markers (alpha actinin-cardiomyocyte marker; 1/100 dilution) in the ventricular infarct zone of LAD surgery mice. Scale bars: 50 microns.

    5 um sections of paraffin or OCT embedded hearts were utilized for Massons Trichrome Staining and immunofluorescence. For SNAIL1 immunofluorescence, antigen retrieval was done in Nuclear Decloaker solution and TSA plus kit was used to amplify SNAIL1 signal.

  • All lanes : Anti-Sarcomeric Alpha Actinin antibody [EA-53] (ab9465) at 1/250 dilution

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : 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: 103 kDa
    Observed band size: 103 kDa
    Additional bands at: 25 kDa, 57 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 90 seconds
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue staining Sarcomeric Alpha Actinin with ab9465.

References

This product has been referenced in:
  • Fu H  et al. Dioxin and AHR impairs mesoderm gene expression and cardiac differentiation in human embryonic stem cells. Sci Total Environ 651:1038-1046 (2019). Read more (PubMed: 30266049) »
  • Redd MA  et al. Patterned human microvascular grafts enable rapid vascularization and increase perfusion in infarcted rat hearts. Nat Commun 10:584 (2019). Read more (PubMed: 30718840) »
See all 85 Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews

Application
IHC - Wholemount
Sample
Rat Tissue (skeletal muscle)
Specification
skeletal muscle

Abcam user community

Verified customer

Submitted Mar 29 2019

Application
Immunohistochemistry (Frozen sections)
Sample
Sheep Tissue sections (Muscle)
Permeabilization
Yes - 0,2 % Triton-X in PBS
Specification
Muscle
Fixative
Methanol:Acetone 1:1

Frau. Kassandra Walluks

Verified customer

Submitted Apr 24 2018

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Heart)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization
Yes - 0.2% Triton X-100
Specification
Heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 26 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Cardiac cells)
Specification
Cardiac cells
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 16 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Heart)
Antigen retrieval step
None
Permeabilization
Yes - Triton x-100, 0.01%
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 05 2015

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (heart)
Permeabilization
Yes - Triton x-100, 0.01%
Specification
heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 17 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Rat 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

Dr. Shyam Sundar Nandi

Verified customer

Submitted Nov 25 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Sample
Rat Cell (Heart)
Specification
Heart
Permeabilization
Yes - Triton x-100, 0.01%
Fixative
Paraformaldehyde

Dr. Shyam Sundar Nandi

Verified customer

Submitted Nov 20 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (cardiomyocytes differentiated from mouse ES cells)
Specification
cardiomyocytes differentiated from mouse ES cells
Fixative
Paraformaldehyde
Permeabilization
Yes - triton-X100
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Feb 18 2010

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (mouse brain parasagittal sections)
Specification
mouse brain parasagittal sections
Fixative
Acetone
Permeabilization
Yes - Ventana protocol

Abcam user community

Verified customer

Submitted Jun 19 2007

1-10 of 11 Abreviews

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