Name: Anti-ACTN3 antibody [EP2531Y]
Brand: RabMAb
SKU: ab68204
Rating: 5 (4 reviews)

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Western blot - 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

Alexa Fluor® 488 Carrier Free

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.

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

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)

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.

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

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) 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)

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)

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)

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)

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)

Flow cytometry protocols
Immunohistochemistry protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

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

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

Abcam user community

Verified customer

Submitted Aug 22 2016

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

Satu Koskinen

Verified customer

Submitted Aug 01 2016

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

Abcam user community

Verified customer

Submitted Jul 25 2016

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

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

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.

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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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Immunizing Peptide (Blocking)

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (4)

Customer reviews and Q&As

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

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

Western blot abreview for Anti-ACTN3 antibody [EP2531Y]

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