Anti-alpha smooth muscle Actin antibody (ab5694)

Overview

  • Product nameAnti-alpha smooth muscle Actin antibody
    See all alpha smooth muscle Actin primary antibodies
  • Description
    Rabbit polyclonal to alpha smooth muscle Actin
  • SpecificityThis antibody stains smooth muscle cells in vessel walls, gut wall, and myometrium. Myoepithelial cells in breast and salivary gland are also stained. It reacts with tumors arising from smooth muscles and myoepithelial cells. The other actins, such as beta- and gamma-cytoplasmic, striated muscle, and myocardium are not stained by this antibody.
  • Tested applicationsSuitable for: IHC-FoFr, ICC, ICC/IF, WB, ELISA, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Guinea pig, Cow, Dog, Human, Pig
  • Immunogen

    Synthetic peptide corresponding to Human alpha smooth muscle Actin. This antibody was raised against a synthetic peptide corresponding to N-terminus of actin from human smooth muscle.

  • General notes

    Actins are highly conserved proteins expressed in all eucaryotic cells. Actin filaments form part of the cytoskeleton and play essential roles in regulating cell shape and movement. Six distinct actin isotypes have been identified in mammalian cells. Each is encoded by a separated gene and is expressed in a developmentally regulated and tissue-specific manner, alpha and beta cytoplasmic actins are expressed in a wide variety of cells; whereas, expression of alpha skeletal, alpha cardiac, alpha vascular, and gamma enteric actins are more restricted to specialized muscle cell type. Smooth muscle alpha actin is of further interest because it is one of a few genes whose expression is relatively restricted to vascular smooth muscle cells. Further more, expression of smooth muscle alpha actin is regulated by hormones, cell proliferation , and altered by pathological conditions including oncogenic transformation and atherosclerosis.

Properties

Applications

Our Abpromise guarantee covers the use of ab5694 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
IHC-FoFr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
ICC/IF 1/100.
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 42 kDa.
ELISA Use a concentration of 0.1 - 1 µg/ml.
IHC-P 1/50 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr 1/200. PubMed: 18559614Fix with acetone.

Target

  • FunctionActins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
  • Involvement in diseaseDefects in ACTA2 are the cause of aortic aneurysm familial thoracic type 6 (AAT6) [MIM:611788]. AATs are characterized by permanent dilation of the thoracic aorta usually due to degenerative changes in the aortic wall. They are primarily associated with a characteristic histologic appearance known as 'medial necrosis' or 'Erdheim cystic medial necrosis' in which there is degeneration and fragmentation of elastic fibers, loss of smooth muscle cells, and an accumulation of basophilic ground substance.
  • Sequence similaritiesBelongs to the actin family.
  • Cellular localizationCytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • a actin antibody
    • AAT6 antibody
    • ACTA_HUMAN antibody
    • ACTA2 antibody
    • Actin alpha 2 smooth muscle aorta antibody
    • Actin aortic smooth muscle antibody
    • Actin, aortic smooth muscle antibody
    • ACTSA antibody
    • ACTVS antibody
    • Alpha 2 actin antibody
    • Alpha actin 2 antibody
    • Alpha cardiac actin antibody
    • Alpha-actin-2 antibody
    • Cell growth inhibiting gene 46 protein antibody
    • Cell growth-inhibiting gene 46 protein antibody
    • GIG46 antibody
    • Growth inhibiting gene 46 antibody
    • MYMY5 antibody
    see all

Anti-alpha smooth muscle Actin antibody images

  • ab5694 at 1/500 staining rat myofibroblast cells by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed and blocked with 5% serum prior to incubation with the antibody for 2 hours. A FITC conjugated goat anti-rabbit IgG was used as the secondary. Nuclei were counterstained with propidium iodide.

    See Abreview

  • Lanes 1 - 3 : Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/ml
    Lane 4 :
    Lane 5 :

    Lane 1 : HEK293 cell lysate - overexpressing alpha-Actin
    Lane 2 : 3T3 cell lysate
    Lane 3 : Mouse heart tissue homogenate
    Lane 4 : 3T3 cell lysate
    Lane 5 : Mouse heart tissue homogenate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Fluor 750-conjugated goat anti-rabbit IgG (H+L) at 1/12500 dilution

    Predicted band size : 42 kDa
    Observed band size : 42 kDa

    Incubated with the primary antibody at 4°C overnight.

    Incubated with the secondary antibody at room temperature for 1 hour.

  • ab5694 staining alpha smooth muscle actin in mouse MEF cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 1% Triton X-100 in PBS and blocked with 10% goat serum for 60 minutes at room temperature. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C in 10% goat serum. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal at a dilution of 1/400 was used as the secondary antibody.

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  • ab5694 staining Human fetal heart cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37°C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37°C. A  Alexa Fluor® 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.

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  • ab5694 staining human Induced pluripotent stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 1% Triton X-100 in PBS and blocked with 10% goat serum for 60 minutes at room temperature. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C in 10% goat serum buffer. An undiluted Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

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  • This picture shows formalin-fixed, paraffin embedded mouse intestine and mesentery, the optimal dilution is 1:1600 to 1:3200, incubation overnight at 4oC, counterstained with Hematoxylin.

    This image was kindly supplied as part of the review by JQ Zhang.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling alpha smooth muscle Actin with ab5694 at a dilution of 1/1000. Heat mediated antigen retrieval was performed for 35 minutes followed by cooling for 20 minutes. Sections were incubated with the primary antibody for 1 hour followed by incubation with a biotinylated secondary antibody for 30 minutes then HRP-Streptavidin for 30 minutes. Developed using DAB chromogen substrate (5-10 minutes). Counter stained with hematoxylin.

    Magnification: left - 10X, right - 40X.

  • Immunohistochemistry (Formalin-fixed paraffin-embedded sections) analysis of skeletal muscle tissue (left) incubated with ab5694 at 1/100 at room temperature for 1 hour showing no specific staining. Right - human tonsil tissue secondary only control.

    Heat mediated antigen retrieval was performed for 35 minutes followed by cooling for 20 minutes. A biotinylated secondary antibody was used for 30 minutes followed by incubation with HRP-Streptavidin for 30 minutes. Developed using DAB chromogen substrate (5-10 minutes). Counter stained with hematoxylin. Magnification 10X.

  • Human Leiomyoma stained with ab5694.
  • All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/ml

    Lane 1 : HeLa Nuclear
    Lane 2 : HeLa whole cell
    Lane 3 : A431 cell lysate
    Lane 4 : Jurkat cell lysate
    Lane 5 : HEK293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Alexa Fluor anti-rabbit at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 42 kDa
    Observed band size : 42 kDa
    Additional bands at : 30 kDa,35 kDa,37 kDa,50 kDa,75 kDa. We are unsure as to the identity of these extra bands.
    Please note that ab5694 does not appear to be specific to smooth muscle.
  • Ab5694 positively staining smooth muscle cells in blood vessels and myoepithelial cells in the frozen tissue of cancerous human mammary gland (pink) at 1/100 dilution. Secondary: CY5 conjugated goat anti rabbit (1/100). Co immunostaining of glandular cell cytokeratin can be seen stained by FITC (green). Auto fluorescent erythrocytes that are present within blood vessels are shown (red), whilst the DAPI counter stain may clearly be seen staining nuclei (blue).

    This image is courtesy of an Abreview submitted by on 22 August 2005. We do not have any further information relating to this image.

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  • All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1/500 dilution

    Lane 1 : Rat2 myofibroblasts (untreated before treatment-0 days)
    Lane 2 : Rat2 myofibroblasts (untreated for 5 days)
    Lane 3 : Rat2 myofibroblasts (treated with 1ng/mL TGF beta)
    Lane 4 : Rat2 myofibroblasts (treated with 10ng/mL TGF beta)
    Lane 5 : Positive control (NIH3T3)
    Lane 6 : Negative control (MDA-MB-469 breast carcinoma cells)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Donkey anti rabbit (HRP) at 1/2500 dilution

    Performed under reducing conditions.

    Predicted band size : 42 kDa

    This image is an edited version of an image submitted courtesy of an Abreview on 20 September 2005. We do not have any further information relating to this image.

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  • All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1/1000 dilution

    Lane 1 : Lystates prepared from pig heart tissue from normal control animals
    Lane 2 : Lystates prepared from pig heart tissue from normal control animals
    Lane 3 : Lystates prepared from pig heart tissue from experimental animals
    Lane 4 : Lystates prepared from pig heart tissue from experimental animals

    Lysates/proteins at 4 µg per lane.

    Secondary
    HRP-conjugated goat polyclonal to rabbit IgG at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size : 42 kDa
    Observed band size : 45 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    This image is a courtesy of Mario Torrado

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  • ab5694 staining alpha smooth muscle Actin in human skin tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in Citrate pH 6.0 and then blocked with 10% serum for 1 hour at RT. The primary antibody was diluted 1/300 and incubated with sample in 2% serum for 15 hours at 4°C. A Biotin conjugated goat polyclonal to rabbit IgG was used at dilution at 1/500 as secondary antibody.

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  • ab5694 staining alpha smooth muscle Actin in rat lung tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then permeabilized using 0.5 Triton X-100 for 20 minutes, blocked with 1% BSA for 30 minutes at 20°C and then incubated with ab5694 at a 1/100 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 568 conjugated goat anti-rabbit polyclonal used at a 1/250 dilution.

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References for Anti-alpha smooth muscle Actin antibody (ab5694)

This product has been referenced in:
  • Kanazawa H  et al. Durable Benefits of Cellular Postconditioning: Long-Term Effects of Allogeneic Cardiosphere-Derived Cells Infused After Reperfusion in Pigs with Acute Myocardial Infarction. J Am Heart Assoc 5:N/A (2016). IHC-P ; Pig . Read more (PubMed: 26857066) »
  • Chung SI  et al. Development of a transgenic mouse model of hepatocellular carcinoma with a liver fibrosis background. BMC Gastroenterol 16:13 (2016). IHC-P ; Mouse . Read more (PubMed: 26821924) »

See all 538 Publications for this product

Product Wall

I have checked the website and the datasheets of these three antibodies and it seems that only ab18147 has been tested for cross-reactivity with rabbit. The other two antibodies are not known yet if they recognize rabbit alpha smooth muscle Actin.
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We do not have set expiration dates for our products. I am therefore also not aware that the information on the shelf life was changed. We indeed increased the length of our Abpromise guarantee to six month instead of 120 days.

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I have some information which may be of interest to you in regards to anti-alpha smooth muscle Actin antibody (ab5694).

I have found reports of alpha-SMA expression in endothelial cells. These can be found from the links below:

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Application Western blot
Sample Mouse Tissue lysate - whole (lung)
Gel Running Conditions Non-reduced Denaturing (12%)
Loading amount 30 µg
Specification lung
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Doohee Shim

Verified customer

Submitted May 25 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: pH9, Tris/EDTA based
Permeabilization No
Specification Colon
Blocking step Serum Free Protein Block as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 24°C
Fixative 10% NBF
Username

Ms. Premier Laboratory

Verified customer

Submitted Apr 07 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: pH9, Tris/EDTA based
Permeabilization No
Specification Colon
Blocking step Serum Free Protein Block as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 24°C
Fixative 10% NBF
Username

Ms. Premier Laboratory

Verified customer

Submitted Apr 07 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Rat Placenta)
Antigen retrieval step Enzymatic
Permeabilization No
Specification Rat Placenta
Blocking step Serum as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 1.5%
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 26 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 10 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Mar 04 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 10 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Mar 04 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 10 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Mar 04 2016

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