Anti-alpha smooth muscle Actin 抗体 (ab5694)

Incubated with the primary antibody at 4°C overnight.

Incubated with the secondary antibody at room temperature for 1 hour.

Passive CLARITY technique (PACT)-sorbitol refractive index matching solution (sRIMS) clearing and 3D imaging of virgin and lactating mouse mammary tissue. a PACT-sRIMS tissue clearing and immunostaining protocol and timeline. Three-dimensional confocal imaging of PACT-sRIMS-cleared virgin (b) and lactating (c) mammary glands immunostained with basal cell markers (K5 and smooth muscle actin (stained with ab5694)) and luminal cell markers (K8 and E-cadherin (E-CAD)). Main image shows the maximum intensity projection of the entire image sequence, with thin optical slices (1 μm) and their depth (z value) relative to the first image in the image sequence.

From a paper comparing imaging of intact virgin and lactating mammary glands using 3D imaging of solvent-cleared organs, see deep brain (seeDB), clear unobstructed brain imaging cocktails (CUBIC) and passive clarity technique.

This picture shows formalin-fixed, paraffin embedded mouse intestine and mesentery, the optimal dilution is 1:1600 to 1:3200, incubation overnight at 4^oC, counterstained with Hematoxylin.

This image was kindly supplied as part of the review by JQ Zhang.

Pancreatic vessel imaging in the intact adult mouse pancreas. In adult mouse tissues (12 weeks old), imaging was performed after CLARITY. Three-dimensional projection clarified mouse pancreas with capillary immunostained for α-smooth muscle actin (green). Scale bar, 200 μm.

From a study, that aimed to improve the original CLARITY procedure and developed specific CLARITY protocols for various intact organs.

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Effects of the ROCK1 inhibition on pancreatic cancer cells and cancer-associated fibroblasts

Fluorescence microscopic analysis of fasudil treated, co-cultured pancreatic cancer cells and cancer-associated fibroblasts. Cells were treated with fasudil for 48 hours and then were stained for α-SMA (red), Collagen I (green), and DNA (blue).

Alpha smooth muscle Actin (α-SMA) is detected ising ab5694 in 5% formaldehyde-fixed cells.

(From Figure 3C of Watcott et al)

Representative histology of aortic valve leaflets from aged mice demonstrates changes in pRb cKO AoV

A) Masson’s trichrome showing reduced collagen staining (blue) in leaflet from pRb cKO mouse with aortic regurgitation (AR). B) Movat pentachrome showing more diffuse collage staining (yellow) in fibrosa, but normal proteoglycan staining (blue) in the spongiosa layer of the leaflet from pRb cKO with AR. C) Immunohistochemistry for α-SMA, demonstrating presence of activated myofibroblasts throughout leaflets of pRb cKO mouse with and without AR. Scale bar is 50μm.

Alpha smooth muscle Actin is detected witn ab5694 at 1/1000 dilution.

(From Figure 2 of Freytsis et al)

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.

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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.

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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.

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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.

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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 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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