ACTN2 Polyclonal antibody

ACTN2 Polyclonal Antibody for IF, IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IP, IHC, IF, ELISA

Conjugate

Unconjugated

Publications

16

Reviews (1)

Cat no : 14221-1-AP

Synonyms

actinin, alpha 2, ACTN2, Alpha actinin 2, CMD1AA, F actin cross linking protein

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Validation Data Gallery

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  WB analysis using 14221-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 14221-1-AP (ACTN2 antibody) at dilution of 1:30000 incubated at room temperature for 1.5 hours.

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  WB analysis of C2C12 using 14221-1-AP

  C2C12 cell were subjected to SDS PAGE followed by western blot with 14221-1-AP (ACTN2 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse skeletal muscle using 14221-1-AP

  mouse skeletal muscle tissue were subjected to SDS PAGE followed by western blot with 14221-1-AP (ACTN2 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse lung using 14221-1-AP

  mouse lung tissue were subjected to SDS PAGE followed by western blot with 14221-1-AP (ACTN2 antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 14221-1-AP

  IP Result of anti-ACTN2 (IP:14221-1-AP, 4ug; Detection:14221-1-AP 1:1000) with HeLa cells lysate 1080ug.

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  IHC staining of mouse heart using 14221-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse heart tissue slide using 14221-1-AP (ACTN2 antibody) at dilution of 1:500 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse skeletal muscle using 14221-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue slide using 14221-1-AP (ACTN2 antibody) at dilution of 1:500 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of Human iPSC derived cardiomyocyte using 14221-1-AP

  Human iPSC derived cardiomyocyte in 2D culture, with alpha actinin (14221-1-AP) and DAPI. Image courtesy of Chandan Kadur Nagaraju, Experimental Cardiology, KU Leuven, Belgium.

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  IF Staining of mouse heart using 14221-1-AP

  Immunofluorescent analysis of (4% PFA) fixed mouse heart tissue using 14221-1-AP (ACTN2 antibody) at dilution of 1:400 and Alexa Fluor 488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of C2C12 using 14221-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed C2C12 cells using ACTN2 antibody (14221-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), 594phalloidin (red).

• 

  IF Staining of mouse heart using 14221-1-AP

  Mouse cardiac tissue slice of 300µm thickness labelled with alpha actinin (14221-1-AP). Image courtesy of Chandan Kadur Nagaraju, Experimental Cardiology, KU Leuven, Belgium.

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  IF Staining of NIH/3T3 using 14221-1-AP

  Immunofluorescent analysis of (-20℃ Ethanol) fixed NIH/3T3 cells using 14221-1-AP (ACTN2 antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of C2C12 using 14221-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed C2C12 cells using 14221-1-AP (ACTN2 antibody), at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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Tested Applications

• Positive WB detected in: mouse brain tissue, C2C12 cells, mouse lung tissue, mouse skeletal muscle tissue, mouse heart, mouse kidney, rat skeletal muscle
• Positive IP detected in: HeLa cells
• Positive IHC detected in: mouse heart tissue, mouse skeletal muscle tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: Human iPSC derived cardiomyocyte, mouse heart tissue, C2C12 cells, NIH/3T3 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for IP and 1:500-1:2000 for WB
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF): IF : 1:10-1:100

Sample-dependent, check data in validation data gallery

Published Applications

• WB: See 10 publications below
• IHC: See 2 publications below
• IF: See 8 publications below

Product Information

14221-1-AP targets ACTN2 in WB, IP, IHC, IF, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: ACTN2 fusion protein Ag5459
• Full Name: actinin, alpha 2
• Calculated molecular weight: 104 kDa
• Observed molecular weight: 103 kDa
• GenBank accession number: BC051770
• Gene symbol: ACTN2
• Gene ID (NCBI): 88
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Alpha actinin 2 (ACTN2) belongs to the alpha-actinin family and is expressed in both skeletal and cardiac muscles and functions to anchor myofibrillar actin thin filaments and titin to Z-discs (PMID: 30701273). ACTN2 is an actin-binding protein with multiple roles in different cell types. In nonmuscle cells, the cytoskeletal isoform is found along microfilament bundles and adherens-type junctions, where it is involved in binding actin to the membrane. In contrast, skeletal, cardiac, and smooth muscle isoforms are localized to the Z disc and analogous dense bodies, where they help anchor the myofibrillar actin filaments. Mutations in ACTN2 are associated with hypertrophic cardiomyopathy, as well as dilated cardiomyopathy and endocardial fibroelastosis (PMID: 20022194, 14567970).

Publications

Species	Application	Title
mouse	WB,IF	Nat Commun; Microskeletal stiffness promotes aortic aneurysm by sustaining pathological vascular smooth muscle cell mechanosensation via Piezo1.; Authors - Weiyi Qian
rat	WB	Basic Res Cardiol; Alkaline nucleoplasm facilitates contractile gene expression in the mammalian heart.; Authors - Alzbeta Hulikova
human	IF	Dev Cell; Essential role of MESP1-RING1A complex in cardiac differentiation; Authors - Qianqian Liang
human	IF	Front Cell Dev Biol; Targeting of CAT and VCAM1 as Novel Therapeutic Targets for DMD Cardiomyopathy.; Authors - Bin Li
human	WB,IF	Front Cell Dev Biol; Incomplete Assembly of the Dystrophin-Associated Protein Complex in 2D and 3D-Cultured Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.; Authors - Guillaume Gilbert
mouse	WB,IF	J Mol Cell Cardiol; LncRNA-Mhrt regulates cardiac hypertrophy by modulating the miR-145a-5p/KLF4/myocardin axis.; Authors - Yao Xu

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.

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FH

Rebecca (Verified Customer) (09-21-2022)

20ug of protein was loaded. Transference at 4ºC and 90V, for 2h. Antibody incubation ON at 4ºC.

• Applications: Western Blot
• Primary Antibody Dilution: 1:10000
• Cell Tissue Type: iPSC-CM

ACTN2 Antibody Western Blot validation (1:10000 dilution) in iPSC-CM (Cat no:14221-1-AP)