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Western Blot (WB) (Figure 1. Western blot analysis of ACTN3 using anti-ACTN3 antibody (M02693). Electrophoresis was performed on a 10% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat skeletal muscle tissue, Lane 2: mouse skeletal muscle tissue. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-ACTN3 antigen affinity purified monoclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. )

Mouse anti-Mouse, Rat ACTN3 Monoclonal Antibody | anti-ACTN3 antibody

Anti-ACTN3 Antibody (monoclonal, 9B5)

Gene Names
ACTN3; ACTN3D
Reactivity
Mouse, Rat
Applications
Western Blot, Immunohistochemistry
Purity
Immunogen Affinity Purified
Synonyms
ACTN3; Monoclonal Antibody; Anti-ACTN3 Antibody (monoclonal; 9B5); Alpha-actinin-3; Alpha-actinin skeletal muscle isoform 3; F-actin cross-linking protein; Actinin alpha 3 (gene/pseudogene); anti-ACTN3 antibody
Ordering
For Research Use Only!
Host
Mouse
Reactivity
Mouse, Rat
Clonality
Monoclonal
Isotype
IgG1
Specificity
No cross reactivity with other proteins.
Purity/Purification
Immunogen Affinity Purified
Form/Format
Lyophilized
Sequence Length
901
Applicable Applications for anti-ACTN3 antibody
Western Blot (WB), Immunohistochemistry (IHC) Paraffin
Application Notes
WB: 0.1-0.5 mug/ml
IHC-P: 0.5-1 mug/ml
Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections.
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human ACTN3 (574-617aa EADRERGAIMGIQGEIQKICQTYGLRPCSTNPYITLSPQDINT K), different from the related mouse sequence by five amino acids.
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Relevant Detection Systems
It is recommended to use an Enhanced Chemiluminescent Kit with anti-Rabbit IgG (MBS176460) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (MBS176453) for IHC-P.
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time.
Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of ACTN3 using anti-ACTN3 antibody (M02693). Electrophoresis was performed on a 10% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat skeletal muscle tissue, Lane 2: mouse skeletal muscle tissue. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-ACTN3 antigen affinity purified monoclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. )

Western Blot (WB) (Figure 1. Western blot analysis of ACTN3 using anti-ACTN3 antibody (M02693). Electrophoresis was performed on a 10% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat skeletal muscle tissue, Lane 2: mouse skeletal muscle tissue. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-ACTN3 antigen affinity purified monoclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. )
Related Product Information for anti-ACTN3 antibody
Description: Mouse IgG monoclonal antibody for ACTN3 detection. Tested with WB, IHC-P in Mouse; Rat.
Background: Alpha-actinin-3, also known as alpha-actinin skeletal muscle isoform 3 or F-actin cross-linking protein, is a protein that in humans is encoded by the ACTN3 gene. This gene encodes a member of the alpha-actin binding protein gene family. The encoded protein is primarily expressed in skeletal muscle and functions as a structural component of sarcomeric Z line. This protein is involved in crosslinking actin containing thin filaments. An allelic polymorphism in this gene results in both coding and non-coding variants; the reference genome represents the coding allele. The non-functional allele of this gene is associated with elite athlete status.
References
1. Papadimitriou ID, Papadopoulos C, Kouvatsi A, Triantaphyllidis C (April 2008). "The ACTN3 gene in elite Greek track and field athletes". Int J Sports Med 29 (4): 352-5. 2. Yang N, MacArthur DG, Gulbin JP, Hahn AG, Beggs AH, Easteal S, North K (September 2003). "ACTN3 genotype is associated with human elite athletic performance". Am. J. Hum. Genet. 73 (3): 627-31.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
89
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
103,241 Da
NCBI Official Full Name
alpha-actinin-3 isoform 1
NCBI Official Synonym Full Names
actinin alpha 3 (gene/pseudogene)
NCBI Official Symbol
ACTN3
NCBI Official Synonym Symbols
ACTN3D
NCBI Protein Information
alpha-actinin-3
UniProt Protein Name
Alpha-actinin-3
Protein Family
UniProt Gene Name
ACTN3

NCBI Description

This gene encodes a member of the alpha-actin binding protein gene family. The encoded protein is primarily expressed in skeletal muscle and functions as a structural component of sarcomeric Z line. This protein is involved in crosslinking actin containing thin filaments. An allelic polymorphism in this gene results in both coding and non-coding variants; the reference genome represents the coding allele. The non-functional allele of this gene is associated with elite athlete status. [provided by RefSeq, Feb 2014]

Uniprot Description

F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein.

Research Articles on ACTN3

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Product Notes

The ACTN3 actn3 (Catalog #AAA1752228) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-ACTN3 Antibody (monoclonal, 9B5) reacts with Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's ACTN3 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC) Paraffin. WB: 0.1-0.5 mug/ml IHC-P: 0.5-1 mug/ml Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. Researchers should empirically determine the suitability of the ACTN3 actn3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "ACTN3, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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