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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microfilaments Actin etc Actin Crosslinking
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RecombinantRabMAb

Recombinant Anti-ACTN3 antibody [EP2531Y] (ab68204)

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

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Western blot - Anti-ACTN3 antibody [EP2531Y] (ab68204)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACTN3 antibody [EP2531Y] (ab68204)
  • Flow Cytometry (Intracellular) - Anti-ACTN3 antibody [EP2531Y] (ab68204)
  • Western blot - Anti-ACTN3 antibody [EP2531Y] (ab68204)
  • Western blot - Anti-ACTN3 antibody [EP2531Y] (ab68204)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACTN3 antibody [EP2531Y] (ab68204)
  • Anti-ACTN3 antibody [EP2531Y] (ab68204)

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: Flow Cyt (Intra), WB, IHC-P
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free

You may also be interested in

Protein
Recombinant Human ACTN3 protein (ab127135)
Primary
Product image
Anti-beta Sarcoglycan antibody (ab135954)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

View more associated products

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: Flow Cyt (Intra), WB, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab127135)

  • Positive control

    • WB: Human skeletal muscle, Mouse skeletal muscle, and Rat skeletal muscle lysates. IHC-P: human skeletal muscle tissue. Flow Cyt (intra): 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP2531Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Crosslinking
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Binding Proteins

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-ACTN3 antibody [EP2531Y] (ab210420)
    • Anti-ACTN3 antibody [EP2531Y] - BSA and Azide free (ab247376)
  • Immunizing Peptide (Blocking)

    • Recombinant Human ACTN3 protein (ab127135)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human ACTN3 protein (ab127135)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/30.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB (1)
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.

Notes
Flow Cyt (Intra)
1/30.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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.

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.
  • Target information above from: UniProt accession Q08043 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 89 Human
    • Entrez Gene: 11474 Mouse
    • Entrez Gene: 171009 Rat
    • Omim: 102574 Human
    • SwissProt: Q08043 Human
    • SwissProt: O88990 Mouse
    • Unigene: 654432 Human
    • Unigene: 737862 Human
    • Unigene: 5316 Mouse
    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 (Intracellular) - Anti-ACTN3 antibody [EP2531Y] (ab68204)
    Flow Cytometry (Intracellular) - Anti-ACTN3 antibody [EP2531Y] (ab68204)

    Intracellular 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.

  • Anti-ACTN3 antibody [EP2531Y] (ab68204)
    Anti-ACTN3 antibody [EP2531Y] (ab68204)

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (4)

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

ab68204 has been referenced in 4 publications.

  • Wyckelsma VL  et al. Loss of a-actinin-3 during human evolution provides superior cold resilience and muscle heat generation. Am J Hum Genet 108:446-457 (2021). PubMed: 33600773
  • Seo JY  et al. Maintenance of type 2 glycolytic myofibers with age by Mib1-Actn3 axis. Nat Commun 12:1294 (2021). PubMed: 33637766
  • Lornage X  et al. ACTN2 mutations cause "Multiple structured Core Disease" (MsCD). Acta Neuropathol 137:501-519 (2019). PubMed: 30701273
  • Koskinen SOA  et al. Human skeletal muscle type 1 fibre distribution and response of stress-sensing proteins along the titin molecule after submaximal exhaustive exercise. Histochem Cell Biol 148:545-555 (2017). PubMed: 28712031

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
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
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
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
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

Abcam community

Verified customer

Asked on Mar 11 2013

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

Abcam Scientific Support

Answered on Mar 11 2013

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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