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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.
Our Abpromise guarantee covers the use of ab5694 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-FoFr||Use at an assay dependent concentration.|
|ICC||Use at an assay dependent concentration.|
|WB||Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 42 kDa.|
|ELISA||Use a concentration of 0.1 - 1 µg/ml.|
|IHC-P||1/50 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IHC-Fr||1/200. PubMed: 18559614Fix with acetone.|
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.
Incubated with the primary antibody at 4°C overnight.
Incubated with the secondary antibody at room temperature for 1 hour.
ab5694 staining alpha smooth muscle actin in mouse MEF cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 1% Triton X-100 in PBS and blocked with 10% goat serum for 60 minutes at room temperature. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C in 10% goat serum. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal at a dilution of 1/400 was used as the secondary antibody.
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.
ab5694 staining human Induced pluripotent stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 1% Triton X-100 in PBS and blocked with 10% goat serum for 60 minutes at room temperature. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C in 10% goat serum buffer. An undiluted Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.
This picture shows formalin-fixed, paraffin embedded mouse intestine and mesentery, the optimal dilution is 1:1600 to 1:3200, incubation overnight at 4oC, counterstained with Hematoxylin.
This image was kindly supplied as part of the review by JQ Zhang.
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.
Magnification: left - 10X, right - 40X.
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.
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.
This image is a courtesy of Mario Torrado
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.
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