Recombinant
RabMAb

Recombinant Anti-ACTN3 antibody [EP2531Y] (ab68204)

Reviews (4)Q&A (1)References (1)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2531Y] to ACTN3
  • Suitable for: WB, IHC-P, Flow Cyt
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ACTN3 antibody [EP2531Y]
    See all ACTN3 primary antibodies

  • Description

    Rabbit monoclonal [EP2531Y] to ACTN3

  • Host species

    Rabbit

  • Specificity

    The mouse and  rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cytmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human ACTN3 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Human skeletal muscle, Mouse skeletal muscle, and Rat skeletal muscle lysates. IHC-P: human skeletal muscle tissue. Flow Cyt: L6 cells.

  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab68204 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 1/5000 - 1/10000. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).Can be blocked with Recombinant Human ACTN3 protein (ab127135).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

The mouse and  rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

See IHC antigen retrieval protocols.
Flow Cyt 1/30.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
  • Application notes
    Is unsuitable for IP.

    • 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 only in a subset of type 2 skeletal muscle fibers.

    • 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.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • Actinin alpha 3 antibody
      • actinin, alpha antibody
      • ACTN 3 antibody
      • ACTN3 antibody
      • ACTN3_HUMAN antibody
      • Alpha actinin skeletal muscle antibody
      • Alpha actinin skeletal muscle isoform 3 antibody
      • Alpha-Actinin 3 antibody
      • Alpha-actinin skeletal muscle isoform 3 antibody
      • Alpha-actinin-3 antibody
      • F actin cross linking protein antibody
      • F-actin cross-linking protein antibody
      • MGC117002 antibody
      • MGC117005 antibody
      • OTTHUMP00000219049 antibody
      • OTTHUMP00000219054 antibody
      see all

    Images

    • Western blot - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      Western blot - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      All lanes : Anti-ACTN3 antibody [EP2531Y] (ab68204) at 1/20000 dilution (Purified)

      Lane 1 : Mouse skeletal muscle lysates
      Lane 2 : Rat skeletal muscle lysates

      Lysates/proteins at 15 µg per lane.

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

      Predicted band size: 103 kDa
      Observed band size: 100 kDa
      why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACTN3 antibody [EP2531Y] (ab68204)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue sections labeling ACTN3 with purified ab68204 at 1/250 dilution (0.97 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    • Flow Cytometry - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      Flow Cytometry - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      Flow Cytometry analysis of L6 (Rat skeletal muscle myoblast) cells labeling ACTN3 with purified ab68204 at 1/30 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    • Western blot - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      Western blot - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      All lanes : Anti-ACTN3 antibody [EP2531Y] (ab68204) at 1/5000 dilution (Purified)

      Lane 1 : Human skeletal muscle lysates at 15 µg


      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 103 kDa
      Observed band size: 100 kDa why is the actual band size different from the predicted?

    • Western blot - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      Western blot - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      Anti-ACTN3 antibody [EP2531Y] (ab68204) at 1/10000 dilution ((unpurified)) + human skeletal muscle lysate at 10 µg

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 103 kDa
      Observed band size: 103 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACTN3 antibody [EP2531Y] (ab68204)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACTN3 antibody [EP2531Y] (ab68204)

      Immunohistochemical analysis of paraffin-embedded human muscle tissue using unpurified ab68204 at 1/100-1/250 dilution.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    References (1)

    Publishing research using ab68204? Please let us know so that we can cite the reference in this datasheet.

    ab68204 has been referenced in 1 publication.

    • Lornage X  et al. ACTN2 mutations cause "Multiple structured Core Disease" (MsCD). Acta Neuropathol 137:501-519 (2019). PubMed: 30701273

    Customer reviews and Q&As

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    1-5 of 5 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-ACTN3 antibody [EP2531Y]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Heart)
    Permeabilization
    Yes - Tween-20
    Specification
    Heart
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 22 2016

    Immunohistochemistry (Frozen sections) abreview for Anti-ACTN3 antibody [EP2531Y]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Cell (skeletal muscle/frozen sections, human, rat and mo)
    Permeabilization
    Yes - 0,1 % Triton X-100 in PBS
    Specification
    skeletal muscle/frozen sections, human, rat and mo
    Blocking step
    Serum as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Histofix/Histolab/Sweden
    Read More

    Satu Koskinen

    Verified customer

    Submitted Aug 01 2016

    Western blot abreview for Anti-ACTN3 antibody [EP2531Y]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Skeletal Muscle)
    Gel Running Conditions
    Reduced Denaturing (4%-12%)
    Loading amount
    10 µg
    Specification
    Skeletal Muscle
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 25 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-ACTN3 antibody [EP2531Y]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Human pluripotent stem cell derived cardiomyocyte)
    Permeabilization
    Yes - saponin
    Specification
    Human pluripotent stem cell derived cardiomyocyte
    Blocking step
    Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 21 2015

    Question

    Hello,
    Do you know if the Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330) can be used a reliable positive control for the Anti-ACTN3 [EP2531Y] (ab68204) antibody in western blotting?
    Thank you

    Read More
    Answer

    Thank you for contacting us.
    Skeletal Muscle (Human) Tissue Lysate is recommended as a positive control for ab68204. Both products are recommended for Western Blotting, and are guaranteed to work in this application.
    If you had any problems using these products in the applications and species stated on our datasheet, please do not hesitate to contact us. Our Abpromise guarantees they should work in these conditions, and if they didn’t, we would replace or refund them, if reported within 6 months after purchasing.
    I hope this helps. Please do not hesitate to contact us for further assistance.

    Read More

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