Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)

Overview

  • Product name
    Anti-alpha smooth muscle Actin antibody [1A4]
    See all alpha smooth muscle Actin primary antibodies
  • Description
    Mouse monoclonal [1A4] to alpha smooth muscle Actin
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, ELISA, IHC-P, IHC-Fr, WB, Flow Cyt, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Human, Pig
    Predicted to work with: Cow, Mammal, Baboon
  • Immunogen

    Synthetic peptide corresponding to Human alpha smooth muscle Actin (N terminal).
    Database link: P62736

  • Positive control
    • WB: HeLa whole cell lysate and nuclear lysate, Hek293 whole cell lysate. Flow cytometry: SV40LT-SMC. IHC-P: FFPE Human breast ductal carcinoma. ICC-IF: SV40LT-SMC cells.
  • General notes

    This antibody reacts with smooth muscle cells of blood vessels and parenchymal tissue of intestine, testis and ovary.

    This antibody clone [1A4] is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab7817 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
ICC/IF Use a concentration of 1 - 5 µg/ml.
ELISA 1/200 - 1/1000.
IHC-P Use a concentration of 0.1 - 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
WB 1/100 - 1/300.
Flow Cyt Use 0.1µg for 106 cells.
ICC Use at an assay dependent concentration.

Target

  • Function
    Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
  • Involvement in disease
    Defects 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 similarities
    Belongs to the actin family.
  • Cellular localization
    Cytoplasm > 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 sma 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

Images

  • IHC image of alpha smooth muscle actin staining in a human breast ductal carcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7817, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Ab7817 staining alpha smooth muscle actin in Mouse intestine tissue by Immunohistochemistry-Immunofluorescence. Tissue was fixed with formaldehyde and blocked with 100% Cas-block for 30 minutes at room temperature; antigen retrieval was performed by heat mediated citrate buffer, pH6. The sample was incubated with primary antibody at 1/200 dilution for 16 hours at 4°C. An Alexa Fluor® 488 Goat anti-mouse IgG was used as the secondary antibody at 1/400 dilution. Autofluorescence was blocked with 0.1% Sudan Black in 70% ethanol for 10 minutes at room temperature after antigen retrieval, and followed with 3X wash with PBS-T after antigen retrieval. Image was taken with confocal microscope.

    See Abreview

  • ab7817 stained in SV40LT-SMC cells. The cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab7817 at 5µg/ml overnight at +4°C. The secondary antibody was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1/200 dilution (Samples were incubated with primary antibody for 16 hours at 4ºC with TBS Tween 0,5% Lait 5% NaN3.) + Mouse intestinal tissue at 14 µg with 5% milk for 1 hour at 25ºC.

    Secondary
    Goat anti-Mouse IgG (H+L) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 1 second

    See Abreview

  • Davis FM et al investigates the differentiation potential of mammary stem cells in adults. Ab7817 staining smooth muscle actin in EYFP+ cells from R26[CA]30EYFP mouse at 1:200 dilution. 

  • All lanes : Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : A431 (Human) Whole Cell Lysate
    Lane 4 : Jurkat (Human) Whole Cell Lysate
    Lane 5 : HEK293 (Human) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 30 seconds
  • ab7817 staining alpha smooth muscle Actin (green) in Mouse primary colon myofibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone and blocked with 5% BSA for 30 hours at 25°C. Samples were incubated with primary antibody (1/100 in PBS + 5% BSA) for 2 hours at 25°C. Donkey Anti-Mouse IgG H&L (DyLight® 488) (ab96875) (1/1000) was used as the secondary antibody. Costained with ab92547, Rabbit anti-Vimentin (red).

    See Abreview

  • ab7817 staining alpha smooth muscle Actin in human IMR-90 (Human Lung Fibroblast Cell Line) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% TritonX-100 and blocked with 100% Cad-Block for 30 minutes at room temperature. Samples were incubated with primary antibody 1/200 in antibody diluent buffer for 16 hours at 4°C. An Alexa Fluor® 488-conjugated polyclonal Goat anti-mouse IgG, dilution 1/400, was used as secondary antibody.

    See Abreview

  • ab7817 staining alpha smooth muscle Actin in mouse heart cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with TritonX-100 and blocked with 5% BSA for 30 minutes at room temperature. Samples were incubated with primary antibody 1/100 in blocking buffer for 2 hours. An Alexa Fluor® 488-conjugated Donkey monoclonal to mouse IgG, dilution 1/200, was used as secondary antibody.

    See Abreview

  • ab7817 staining alpha smooth muscle actin in Human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and permeabilized with wash buffer with tween; antigen retrieval was by heat mediation in Tris-EDTA buffer, pH 9.0. Samples were incubated with primary antibody (1/100 in blocking buffer) for 30 minutes at 20°C. A HRP-conjugated Goat anti-mouse IgG polyclonal (undiluted) was used as the secondary antibody.

    See Abreview

  • ab7817 staining alpha smooth muscle Actin in human brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, cut into 20 micron slices, permeablized with 0.1 M PBS with 1% Triton X and blocked with 10% serum for 60 minutes at 24°C. The sample was incubated with primary antibody (1/200 in 0.1M PBST with 10% donkey serum) at 4°C for 24 hours. An Alexa Fluor® 568-conjugated donkey monoclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing SV40LT-SMC cells stained with ab7817 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab7817, 0.1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113) at 1/2000 dilution for 30 min at 22°C

    Isotype control antibody (black line) was mouse IgG2a [18C8BC7AD10] (ab170191) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

  • Immunohistochemical analysis of mouse aorta (A) or skin (B) tissue, staining alpha smooth muscle Actin with ab7817.

    Tissue was fixed with 10% Neutral Buffered Formalin and blocked with 1% serum for 45 minutes 21°C; antigen retrieval was by enzymatic method in 0.0001% Trypsin-CaCl. Samples were incubated with primary antibody (4 µg/ml in 0.3% Triton X-100 in PBS) for 1 hour at 21°C. A biotin-conjugated horse anti-mouse polyclonal IgG (1/50) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Duchalais E  et al. Colorectal Cancer Cells Adhere to and Migrate Along the Neurons of the Enteric Nervous System. Cell Mol Gastroenterol Hepatol 5:31-49 (2018). Read more (PubMed: 29188232) »
  • Charrier EE  et al. Control of cell morphology and differentiation by substrates with independently tunable elasticity and viscous dissipation. Nat Commun 9:449 (2018). Read more (PubMed: 29386514) »

See all 219 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (tumor)
Permeabilization
No
Specification
tumor
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted May 31 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (skeletal muscle)
Permeabilization
No
Specification
skeletal muscle
Fixative
none
Username

Abcam user community

Verified customer

Submitted May 23 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ab94674 100X Citrate Buffer pH 6.0
Permeabilization
Yes - 0.05% tween 20
Specification
colon
Blocking step
Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
Fixative
10%NBF
Username

Mr. David Ivancic

Verified customer

Submitted May 04 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Dog Tissue sections (artery)
Permeabilization
No
Specification
artery
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 30 2018

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 (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Feb 05 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Feb 05 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (breast tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
breast tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Feb 05 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 30 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (Cancer Cell lines)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
10 µg
Specification
Cancer Cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Dec 11 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Vascular smooth muscle cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Vascular smooth muscle cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Sep 11 2017

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