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

Overview

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.
ICC Use at an assay dependent concentration.

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 [1A4] 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 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 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.

  • 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
    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 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 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 Alexa Fluor® 488 goat anti-mouse IgG (H&L) (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.

  • All lanes : Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1/200 dilution

    Lane 1 : HeLa Nuclear
    Lane 2 : HeLa whole cell lysate
    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-mouse at 1/5000 dilution

    Performed under reducing conditions.

    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
    Fluorescence detection of secondary antibody.
  • 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. ab96875, a donkey anti-mouse DyLight®488 (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 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 (red) in Human lung adenocarcinoma cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehye, permeabilized with 0.1% Triton X-100 and blocked with 3% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/400 in 1x TBS) for 3 hours at 25°C. An Alexa Fluor®633-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were counterstained with Hoeches 33258 (blue).

    See Abreview

  • 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.

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  • Cannabinoid receptor (CB1) (green) and α-smooth muscle actin immunoreactivity (red) in a cross-section of rat middle cerebral artery. Rat middle cerebral artery sections were incubated for 12 hours with a 1:300 dilution of anti-CB1 polyclonal antibody raised in rabbits and a 1:100 dilution of anti-α-smooth muscle actin monoclonal antibody raised in mice. The secondary antibodies were Alexa-488-labeled goat anti-rabbit at a 1:500 dilution and Cy3-labeled goat anti-mouse at a 1:1,000 dilution. This picture was submitted by David Rademacher whose review of the antibody is in the review section of this datasheet.

  • Ab7817 positively staining paraformaldehyde fixed porcine valvular interstitial cells (VICs) at 1/75. Staining was carried out in conjunction with goat anti mouse Alexa Fluor® 488 (1/400).

    Ab7817 was used to detect myofibroblsats among a heterogeneous interstitial cell type. The image shows heparin treated VICs labelled for alpha actin (green), and counterstained with propidium iodide (red).

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

    See Abreview

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

This product has been referenced in:
  • Caiazzo M  et al. Defined three-dimensional microenvironments boost induction of pluripotency. Nat Mater N/A:N/A (2016). IF ; Mouse . Read more (PubMed: 26752655) »
  • Izumi D  et al. CXCL12/CXCR4 activation by cancer-associated fibroblasts promotes integrin ß1 clustering and invasiveness in gastric cancer. Int J Cancer 138:1207-19 (2016). WB, IHC ; Human . Read more (PubMed: 26414794) »

See all 88 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (Carotid Artery)
Antigen retrieval step None
Permeabilization No
Specification Carotid Artery
Blocking step Dako Protein Block with casein (Cat X0909) as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: 21°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 06 2017

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (mouse SMC)
Permeabilization Yes - 0.1% TX-100, 0.01% SDS in 1xPBS
Specification mouse SMC
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 01 2017

Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (rat primary fibroblasts)
Permeabilization Yes - 0.5% TX100
Specification rat primary fibroblasts
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Nov 04 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human primary fibroblasts)
Permeabilization Yes - 0.5% TX100
Specification human primary fibroblasts
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Oct 05 2016

Application Western blot
Sample Pig Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 25 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

Submitted Jul 08 2016

Application Western blot
Sample Human Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 14 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

Submitted Jul 08 2016

Application Western blot
Sample Rat Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 14 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

Submitted Jul 08 2016

Application Western blot
Sample Mouse Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 14 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

Submitted Jul 08 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 (muscle section)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization No
Specification muscle section
Blocking step Caseine as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative Bouin Alcohol
Username

Abcam user community

Verified customer

Submitted Mar 16 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Brain)
Permeabilization Yes - 0.1 M PBS with 1% Triton X
Specification Brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative Paraformaldehyde
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Feb 05 2016

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