Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964)

Overview

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab124964 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
WB 1/10000 - 1/50000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).

For unpurified use at 1/1000.

IHC-P 1/1000 - 1/2500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

Flow Cyt 1/10 - 1/100.

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

ICC/IF 1/250 - 1/500.

 

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 [EPR5368] images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovary tissue labelling alpha smooth muscle Actin with unpurified ab124964 at 1/1000 dilution.

  • All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/10000 dilution

    Lane 1 : Human lung lysate
    Lane 2 : A431 cell lysate
    Lane 3 : A549 cell lysate
    Lane 4 : NIH3T3 cell lysate
    Lane 5 : SV40-LT cell lysate
    Lane 6 : C2C12 cell lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique

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


    Exposure time : 10 seconds

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% milk before being incubated with ab124964 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

  • All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/10000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : HEK293 cell lysate
    Lane 3 : U937 cell lysate
    Lane 4 : A431 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 buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/50000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue 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 buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/1000 dilution (unpurified)

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Heart (Mouse) Tissue Lysate
    Lane 3 : Heart (Rat) Tissue Lysate
    Lane 4 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 6 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 7 : SV40LT-SMC (Rat aorta smooth muscle) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 42 kDa
    Additional bands at : 42 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labelling alpha smooth muscl Actin with purified ab124964 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human heart tissue labelling alpha smooth muscle actin with unpurified ab124964 at 1/1000 dilution. Note positive staining on smooth muscle cells but negative on striated muscle cells. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human liver vessels tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human colon smooth muscle tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic carcinoma smooth muscles tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis showing vascular smooth muscle staining in skeletal muscle tissue using  alpha smooth muscle Actin with unpurified ab124964. Note positive staining on smooth muscle cells but negative on striated muscle cells.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach carcinoma smooth muscles tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human tonsil vessels tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunocytochemistry/Immunofluorescence analysis of A673 cells labelling alpha smooth muscle Actin with purified ab124964 at 1/300. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/300) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).

  • Flow cytometry analysis of Jurkat cells labelling alpha smooth muscle Actin with purified ab124964 at 1/30 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964)

This product has been referenced in:

See all 5 Publications for this product

Product Wall

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 post-MI
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 Nov 28 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 20 minute(s) · Concentration: 1.5% · Temperature: 21°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: antigen retrieval
Sample Mouse Tissue sections (Heart-LV at day 0 and day 7 post-MI)
Specification Heart-LV at day 0 and day 7 post-MI
Permeabilization No
Fixative 10% zinc formalin
Username

Dr. Merry Lindsey

Verified customer

Submitted Sep 24 2013

I can confirm that both ab52897 and ab124964 are rabbit IgG monoclonals.

We have several suitable isotype controls available including the following. These have both been tested in IHC-P. I can recommend to review the online datasheets for f...

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