Invitrogen

Skeletal Muscle Actin Monoclonal Antibody (5C5.F8.C7 (alpha-Sr-1))

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

2 Published Figures

7 References

View all (7) Skeletal Muscle Actin antibodies

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Datasheet

Protocols

Questions & Answers

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Skeletal Muscle Actin Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Actin Skeletal Muscle was done on 70% confluent log phase L6 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Actin Skeletal Muscle (5C5.F8.C7 (alpha-Sr-1)) Mouse Monoclonal Antibody (Product # MA5-12542) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L)/IgM (L) Superclonal™ 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 Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Skeletal Muscle Actin Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Actin Skeletal Muscle showing staining in the cytoplasm of paraffin-embedded mouse skeletal muscle tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Actin Skeletal Muscle Mouse Monoclonal Antibody (Product # MA5-12542) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Skeletal Muscle Actin Antibody in Immunohistochemistry (IHC) — Formalin-fixed, paraffin-embedded human skeletal muscle stained with Skeletal Muscle Actin antibody using peroxidase-conjugate and DAB chromogen. Note cytoplasmic staining of skeletal muscle cells.

Skeletal Muscle Actin Antibody in Western Blot (WB) — Figure 3. Actalpha protein localizes to axonal branch points, whereas Actbeta and Actgamma are highly enriched in axonal filopodia and axonal growth cones. (A-C) Immunoblots of total lysates obtained from actin isoform-specific knockdown and control motoneurons probed with specific Actalpha (A), Actbeta (B), and Actgamma (C) antibodies. Calnexin was used as loading control. All three isoforms are detected in cultured motoneurons. (D) shRNA-mediated knockdown leads to a 60% reduction in Actalpha (**, P < 0.004 for n = 4), a 95% reduction in Actbeta (**, P < 0.002 for n = 10), and a 85% reduction in Actgamma (**, P < 0.005 for n = 5) protein levels (one-tailed Mann-Whitney test). Shown are mean +- SEM. (E-G) Motoneurons were stained against Tau and Actalpha (E), Actbeta (F), and Actgamma (G). Actalpha protein is highly enriched in axonal branch points and neurites (E). Actbeta (F) and Actgamma proteins (G) are very abundant in the soma and localize to axonal filopodia and axonal growth cone filopodia. (H-J) Motoneurons were extracted using Triton X-100 to remove G-actin. Cells were fixed and stained with phalloidin and Actalpha (H), Actbeta (I), and Actgamma (J) antibodies. (H) Colocalization of Actalpha with phalloidin shows that this isoform incorporates into F-actin in the axon and particularly in the axonal branch points. (I) Filamentous Actbeta is present predominantly in the axonal growth cone filopodia. (J) Actgamma colocalizes with phalloidin mostly in the soma and axon

Skeletal Muscle Actin Antibody — Figure 3. Actalpha protein localizes to axonal branch points, whereas Actbeta and Actgamma are highly enriched in axonal filopodia and axonal growth cones. (A-C) Immunoblots of total lysates obtained from actin isoform-specific knockdown and control motoneurons probed with specific Actalpha (A), Actbeta (B), and Actgamma (C) antibodies. Calnexin was used as loading control. All three isoforms are detected in cultured motoneurons. (D) shRNA-mediated knockdown leads to a 60% reduction in Actalpha (**, P < 0.004 for n = 4), a 95% reduction in Actbeta (**, P < 0.002 for n = 10), and a 85% reduction in Actgamma (**, P < 0.005 for n = 5) protein levels (one-tailed Mann-Whitney test). Shown are mean +- SEM. (E-G) Motoneurons were stained against Tau and Actalpha (E), Actbeta (F), and Actgamma (G). Actalpha protein is highly enriched in axonal branch points and neurites (E). Actbeta (F) and Actgamma proteins (G) are very abundant in the soma and localize to axonal filopodia and axonal growth cone filopodia. (H-J) Motoneurons were extracted using Triton X-100 to remove G-actin. Cells were fixed and stained with phalloidin and Actalpha (H), Actbeta (I), and Actgamma (J) antibodies. (H) Colocalization of Actalpha with phalloidin shows that this isoform incorporates into F-actin in the axon and particularly in the axonal branch points. (I) Filamentous Actbeta is present predominantly in the axonal growth cone filopodia. (J) Actgamma colocalizes with phalloidin mostly in the soma and axon {KD}

Product Details

MA5-12542

Applications	Tested Dilution	Publications
Western Blot (WB)	-	View 1 publication 1 publication
Immunohistochemistry (IHC)	-	View 5 publications 5 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:200-1:400	-
Immunocytochemistry (ICC/IF)	2 µg/mL	View 1 publication 1 publication

Product Specifications
Species Reactivity	Amphibian, Bovine, Guinea pig, Human, Mouse, Sheep, Rabbit, Rat
Published species	Human, Mouse, Rabbit
Host/Isotype	Mouse / IgM, kappa
Class	Monoclonal
Type	Antibody
Clone	5C5.F8.C7 (alpha-Sr-1)
Immunogen	N-terminal decapeptide
Conjugate	Unconjugated
Form	Liquid
Concentration	0.2 mg/mL
Purification	Ammonium sulfate precipitation
Storage buffer	PBS, pH 7.4, with 0.2% BSA
Contains	0.09% sodium azide
Storage conditions	4° C
RRID	AB_10984949

Product Specific Information

MA5-12542 targets Actin Skeletal Muscle in IF and IHC (P) applications and shows reactivity with Amphibian, Bovine, Guinea Pig, Human, Ovine, Rabbit, and Rat samples.

The MA5-12542 immunogen is n-terminal decapeptide.

Target Information

The product encoded by this gene 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 actins being a major constituent of the contractile apparatus, while beta and gamma actins are involved in the regulation of cell motility. This actin is an alpha actin that is found in skeletal muscle. Mutations in this gene cause nemaline myopathy type 3, congenital myopathy with excess of thin myofilaments, congenital myopathy with cores, and congenital myopathy with fiber-type disproportion, diseases that lead to muscle fiber defects.

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

Product References

In Vivo Analysis of Trypanosoma cruzi Persistence Foci at Single-Cell Resolution.

mBio

Ward AI,Lewis MD,Khan AA,McCann CJ,Francisco AF,Jayawardhana S,Taylor MC,Kelly JM

MA5-12542 was used in Immunohistochemistry to describe an integrated approach involving comprehensive tissue processing, ex vivo imaging, and confocal microscopy, which allowed us to visualize infected host cells in murine tissue with exquisite sensitivity.

Tue Aug 04 00:00:00 EDT 2020

Differential roles of α-, β-, and γ-actin in axon growth and collateral branch formation in motoneurons.

The Journal of cell biology

Moradi M,Sivadasan R,Saal L,Lüningschrör P,Dombert B,Rathod RJ,Dieterich DC,Blum R,Sendtner M

MA5-12542 was used in Immunocytochemistry-immunoflourescence to provide evidence for specific roles of different actin isoforms in spatial regulation of actin dynamics and stability in axons of developing motoneurons.

Mon Mar 06 00:00:00 EST 2017

Differential roles of α-, β-, and γ-actin in axon growth and collateral branch formation in motoneurons.

The Journal of cell biology

Moradi M,Sivadasan R,Saal L,Lüningschrör P,Dombert B,Rathod RJ,Dieterich DC,Blum R,Sendtner M

Mon Mar 06 00:00:00 EST 2017

Epithelial and muscular regionalization of the human developing anorectum.

Anatomical record (Hoboken, N.J. : 2007)

Fritsch H,Aigner F,Ludwikowski B,Reinstadler-Zankl S,Illig R,Urbas D,Schwarzer C,Longato S

MA5-12542 was used in immunohistochemistry to study epithelial and muscular regionalization during development of the human anorectum

Thu Nov 01 00:00:00 EDT 2007

Caveolin-1 expression is associated with plaque formation in hypercholesterolemic rabbits.

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Lin WW,Lin YC,Chang TY,Tsai SH,Ho HC,Chen YT,Yang VC

MA5-12542 was used in immunohistochemistry to study the association of caveolin-1 expression with plaque formation in hypercholesterolemic rabbits

Tue Aug 01 00:00:00 EDT 2006

Spindle cell lipoma in an intramuscular lipoma.

Pathology international

Usta U,Türkmen E,Mizrak B,Yildiz D,Güzel Z

MA5-12542 was used in immunohistochemistry to study a spindle cell lipoma in an intramuscular lipoma

Wed Sep 01 00:00:00 EDT 2004

Clear cell sarcoma of the stomach.

Histopathology

Pauwels P,Debiec-Rychter M,Sciot R,Vlasveld T,den Butter B,Hagemeijer A,Hogendoorn PC

MA5-12542 was used in immunohistochemistry to report on a case of clear cell sarcoma of the stomach

Sun Dec 01 00:00:00 EST 2002

Bioinformatics

Protein Aliases: actin alpha 1; actin, alpha 2, smooth muscle, aorta; Actin, alpha skeletal muscle; Actin, alpha skeletal muscle, intermediate form; actin, alpha, vascular smooth muscle; Actin, aortic smooth muscle; Actin, aortic smooth muscle, intermediate form; alpha actin; alpha actin 2; Alpha-actin-1; Alpha-actin-2; alpha-cardiac actin; alpha-smooth muscle actin; Cell growth-inhibiting gene 46 protein; cytoplasmic beta-actin; nemaline myopathy type 3; smooth muscle alpha-actin; unnamed protein product

Gene Aliases: 0610041G09Rik; a-SMA; AA959943; AAT6; ACTA; Acta-2; ACTA1; ACTA2; Acts; ACTSA; Actsk-1; ACTVS; alphaSMA; ASMA; CFTD; CFTD1; CFTDM; GIG46; MPFD; MYMY5; NEM1; NEM2; NEM3; SHPM; SMalphaA

UniProt ID: (Human) P68133, (Bovine) P68138, (Rabbit) P62740, (Bovine) P62739, (Human) P62736, (Mouse) P68134, (Rat) P68136, (Rat) P62738, (Mouse) P62737

Entrez Gene ID: (Human) 58, (Bovine) 281592, (Rabbit) 100009506, (Sheep) 101107662, (Rabbit) 100009271, (Bovine) 515610, (Human) 59, (Sheep) 101121051, (Guinea pig) 100716933, (Mouse) 11459, (Rat) 29437, (Rat) 81633, (Mouse) 11475, (Guinea pig) 100722917