Invitrogen

Alpha-Smooth Muscle Actin Monoclonal Antibody (1A4), eBioscience™

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated. View Details

4 Published Figures

7 References

View all (27) Alpha-Smooth Muscle Actin antibodies

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Datasheet

Questions & Answers

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Alpha-Smooth Muscle Actin Antibody — Published figure using Alpha-Smooth Muscle Actin monoclonal antibody (Product # 14-9760-82)

Alpha-Smooth Muscle Actin Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of Smooth Muscle Actin was performed using 70% confluent log phase C2C12 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Smooth Muscle Actin Polyclonal Antibody (Product # 14976082) at 1µg/ml in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization to the cytoskeleton. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Alpha-Smooth Muscle Actin Antibody in Immunocytochemistry (ICC) — Immunocytochemistry of methanol-fixed and permeabilized HeLa cells using 1 µg/mL Anti-Alpha-Smooth Muscle Actin Purified, followed by 10 µg/mL F (ab')2 Anti-Mouse IgG eFluor® 570.

Alpha-Smooth Muscle Actin Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Nuclei are stained with DAPI.Immunohistochemistry of formalin-fixed paraffin embedded human infiltrating ductal carcinoma using 1 µg/mL Anti-Alpha-Smooth Muscle Actin Purified, followed by Anti-Mouse IgG Biotin, Streptavidin HRP and DAB visualization (right).Nuclei are counterstained with hematoxylin.

Alpha-Smooth Muscle Actin Antibody in Western Blot (WB) — Western blot was performed using Anti-Alpha-Smooth Muscle Actin Mouse Monoclonal Antibody (Product # 14-9760-80) and a 42kDa band corresponding to Alpha-Smooth Muscle Actin was observed across cell line and tissues tested. Membrane enriched extracts (30 µg lysate) of C2C12 (Lane 1), tissue extracts of Mouse Heart (Lane 2) and Mouse Kidney (Lane 3) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001).The blot was probed with the primary antibody (1:100 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A 25 kDa band (*) corresponding to free IgG was also observed in the tissue extracts.

Alpha-Smooth Muscle Actin Antibody in Relative expression — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell models tested owing to their inherent genetic constitution. Relative expression of Alpha-Smooth Muscle Actin was observed in C2C12 in comparison to Neuro-2a using Anti-Alpha-Smooth Muscle Actin Monoclonal Antibody (Product # 14-9760-82). {RE}

Product Details

Tested Applications

Dilution

Flow Cytometry (Flow)

Assay-Dependent

Immunocytochemistry (ICC)

1 µg/mL

Immunofluorescence (IF)

1 µg/mL

Immunohistochemistry (Frozen) (IHC (F))

1 µg/mL

Immunohistochemistry (Paraffin) (IHC (P))

Assay-Dependent

Immunoprecipitation (IP)

Assay-Dependent

Western Blot (WB)

1:100

Published Applications

Immunofluorescence (IF)

See 4 publications below

Flow Cytometry (Flow)

See 1 publication below

Western Blot (WB)

See 1 publication below

Immunohistochemistry (Frozen) (IHC (F))

See 1 publication below

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Mouse, Rat

Host / Isotype

Mouse / IgG2a, kappa

Class

Monoclonal

Type

Antibody

Clone

1A4

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_2572996

Product Specific Information

Description: The monoclonal antibody 1A4 recognizes human, mouse, and rat alpha-smooth muscle actin. Alpha-smooth muscle actin is a 42 kDa protein and is a major component of the cytoskeletal structural network. This 1A4 antibody is specific for the alpha form of muscle actin only, which is expressed by smooth muscle cells of blood vessels, myofibroblasts, and myoepithelial cells. It is also expressed in the parenchyma and stroma of various tissues. Alpha-smooth muscle actin is useful in the identification of leiomyomas, leiomycosarcomas and pleomorphic adenomas and is used as a prognostic marker for basal cell carcinoma.

Applications Reported: This 1A4 antibody has been reported for use in intracellular staining followed by flow cytometric analysis, immunoprecipitation, western blotting, immunohistochemical staining of frozen tissue sections, immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, microscopy, and immunocytochemistry. (Fluorochrome conjugated 1A4 is recommended for use in intracellular flow cytometry.).

Applications Tested: This 1A4 antibody has been tested by immunohistochemistry of formalin-fixed paraffin embedded human tissue using low or high pH antigen retrieval and can be used at less than or equal to 1 µg/mL. This 1A4 antibody has been tested by immunocytochemistry of formaldehyde or methanol-fixed and permeabilized human cells and can be used at 1 µg/mL. This 1A4 antibody has been tested by western blot analysis of reduced rat and mouse intestine and can be used at less than or equal to 5 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Smooth Muscle Actin belongs to the actin family of proteins, which are highly conserved proteins that play a role in cell motility, structure and integrity. Alpha, beta and gamma actin isoforms have been identified, with alpha actin being a major constituent of the contractile apparatus, while beta and gamma actins are involved in the regulation of cell motility. In particular, smooth muscle actin is an alpha actin that is found in skeletal muscle. Actin exists as a ubiquitous protein involved with filament formation that make up large portions of the cytoskeleton. Actin filaments interact with myosin to assist in muscle contraction as well as aiding in cell motility and cytokinesis. Smooth muscle actin is found on smooth muscle vessel walls, gut wall, myometrium, myoepithelial cells in breast and salivary glands. Defects in the smooth muscle actin gene cause aortic aneurysm familial thoracic type 6. Actin isoforms differ slightly in their N-terminus and the sequences of each are perfectly conserved in higher vertebrates. Alpha-smooth muscle actin is abundant in vascular and visceral smooth muscle cells. In addition, it has also been shown that smooth muscle actin appear in stress fibers of fibroblastic cells during pathological situations involving contractile phenomena such as wound healing and fibrocontractive diseases. Multiple alternatively spliced variants of smooth muscle actin have been identified.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: actin, alpha, vascular smooth muscle; Actin, aortic smooth muscle; Actin, aortic smooth muscle, intermediate form; Alpha Actin 2; Alpha-actin-2; alpha-cardiac actin; alpha-sma; aortic smooth muscle; Cell growth-inhibiting gene 46 protein; smooth muscle alpha-actin

Gene Aliases: 0610041G09Rik; a-SMA; AAT6; ACTA2; ACTSA; ACTVS; alphaSMA; GIG46; MYMY5; SMalphaA

UniProt ID: (Human) P62736, (Rat) P62738, (Mouse) P62737

Entrez Gene ID: (Human) 59, (Rat) 81633, (Mouse) 11475

Molecular Function: actin and actin related protein actin family cytoskeletal protein cytoskeletal protein