This 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.
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.
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.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use at an assay dependent concentration.
Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 42 kDa.
Use a concentration of 0.1 - 1 µg/ml.
1/50 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1/200. PubMed: 18559614Fix with acetone.
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.
Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody (ab5694)This image is courtesy of an Abreview submitted by Dr Jianyuan Chai
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.
Secondary All lanes : 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.
Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody (ab5694)This image is courtesy of an anonymous Abreview
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.
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.
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.
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.
Western blot - Anti-alpha smooth muscle Actin antibody (ab5694)
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 All lanes : 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)This image is courtesy of an Anonymous Abreview
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)Image courtesy of an anonymous Abreview.
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.
Luo L et al. Combination treatment of adipose-derived stem cells and adiponectin attenuates pulmonary arterial hypertension in rats by inhibiting pulmonary arterial smooth muscle cell proliferation and regulating the AMPK/BMP/Smad pathway. Int J Mol Med41:51-60 (2018).
Read more (PubMed: 29115380) »
Freytsis M et al. Conditional deletion of RB1 in the Tie2 lineage leads to aortic valve regurgitation. PLoS One13:e0190623 (2018).
Read more (PubMed: 29304157) »