Anti-alpha smooth muscle Actin antibody [E184] (ab32575)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab32575 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-FrFl Use at an assay dependent concentration. PubMed: 24647450
IHC-Fr 1/100.
WB 1/1000 - 1/5000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human alpha smooth muscle Actin peptide (ab211918).
IHC-P 1/200 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/500.
  • Application notesIs unsuitable for IP.
  • 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 [E184] images

    • Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells labelling alpha smooth muscle Actin (green) with purified ab32575 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by ab150120 AlexaFluor®594 goat anti-mouse secondary (1/1000). Nuclei were counterstained with DAPI (blue).

      For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (ab150077).  

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human smooth muscle tissue labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • All lanes : Anti-alpha smooth muscle Actin antibody [E184] (ab32575) at 1/5000 dilution (purified)

      Lane 1 : A431 whole cell lysate
      Lane 2 : HeLa whole cell lysate
      Lane 3 : MCF-7 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 42 kDa
      Observed band size : 42 kDa
      Blocking and dilution buffer: 5% NFDM/TBST
    • Flow Cytometry analysis of HeLa cells labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/20 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    • All lanes : Anti-alpha smooth muscle Actin antibody [E184] (ab32575) at 1/5000 dilution (purified)

      Lane 1 : C6 whole cell lysate
      Lane 2 : RAW264.7 whole cell lysate
      Lane 3 : PC-12 whole cell lysate
      Lane 4 : NIH/3T3 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 42 kDa
      Observed band size : 42 kDa
      Blocking and dilution buffer: 5% NFDM/TBST
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse smooth muscle tissue labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Overlay histogram showing HeLa cells stained with unpurified ab32575 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab32575, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG; H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Unpurified ab32575 staining alpha smooth muscle actin in human kidney tumour sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 10% serum for 20 minutes at 21°C. Samples were incubated with primary antibody (1/100 in TBS + 2% BSA + 0.02% sodium azide) for 1 hour at 21°C. An undiluted HRP-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.

    References for Anti-alpha smooth muscle Actin antibody [E184] (ab32575)

    This product has been referenced in:
    • Landowski TH  et al. Magnetic Resonance Imaging Identifies Differential Response to Pro-Oxidant Chemotherapy in a Xenograft Model. Transl Oncol 9:228-35 (2016). IHC-P . Read more (PubMed: 27267841) »
    • Soranno DE  et al. Delivery of interleukin-10 via injectable hydrogels improves renal outcomes and reduces systemic inflammation following ischemic acute kidney injury in mice. Am J Physiol Renal Physiol 311:F362-72 (2016). Read more (PubMed: 26962109) »

    See all 34 Publications for this product

    Product Wall

    Application Immunohistochemistry (Frozen sections)
    Sample Mouse Tissue sections (Heart)
    Permeabilization Yes - 0.2% triton TX-100
    Specification Heart
    Blocking step BSA 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 Mar 04 2016

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Mouse Tissue sections (Liver)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
    Permeabilization Yes - Wash Buffer from Dako with Tween
    Specification Liver
    Fixative Paraformaldehyde
    Username

    Mr. Rudolf Jung

    Verified customer

    Submitted Dec 21 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (Skin)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer ph 9,0
    Permeabilization Yes - Wash Buffer from Dako with Tween
    Specification Skin
    Fixative Paraformaldehyde
    Username

    Mr. Rudolf Jung

    Verified customer

    Submitted Dec 21 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Mouse Tissue sections (Lung)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris-EDTA
    Permeabilization Yes - 0.5% tritonX-100
    Specification Lung
    Blocking step invitrogen kit blocking serum as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Sep 09 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: sodium citrate pH 6.0
    Sample Mouse Tissue sections (Skin wound)
    Specification Skin wound
    Permeabilization Yes - 0.05% Tween 20
    Fixative 10% formalin
    Username

    Abcam user community

    Verified customer

    Submitted Oct 27 2014

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Non-reduced Non-Denaturing (Native)
    Sample Mouse Tissue lysate - whole (liver)
    Specification liver
    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 May 05 2014

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Sodium Citrate Buffer pH 6.0
    Sample Mouse Tissue sections (liver)
    Specification liver
    Permeabilization Yes - Sodium Citrate Buffer pH 6.0
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted May 02 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 1.5% · Temperature: 21°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: antigen retrieval
    Sample Mouse Tissue sections (Heart at day 0 and day 7 post-MI)
    Specification Heart at day 0 and day 7 post-MI
    Permeabilization No
    Fixative 10% zinc formalin
    Username

    Dr. Merry Lindsey

    Verified customer

    Submitted Sep 20 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Loading amount 10 µg
    Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris Gel)
    Sample Mouse Tissue lysate - whole (heart)
    Specification heart
    Treatment day 0 and day 7 after myocardial infarction
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Username

    Dr. Merry Lindsey

    Verified customer

    Submitted Sep 20 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step Heat mediated
    Sample Mouse Tissue sections (blood vessels in the soft tissue)
    Specification blood vessels in the soft tissue
    Permeabilization Yes - 0.1% saponin
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Apr 29 2013

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