Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)

Human Leiomyoma stained with ab5694.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)

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.

Western blot - alpha smooth muscle Actin antibody (ab5694)

All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/ml

Lane 1 : HeLa Nuclear
Lane 2 : HeLa whole cell
Lane 3 : A431 cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : HEK293 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
Alexa Fluor anti-rabbit at 1/5000 dilution

Performed under reducing conditions.

Predicted band size : 42 kDa
Observed band size : 42 kDa
Additional bands at : 30 kDa,35 kDa,37 kDa,50 kDa,75 kDa. We are unsure as to the identity of these extra bands.

Please note that ab5694 does not appear to be specific to smooth muscle.

Immunohistochemistry (Frozen sections) - alpha smooth muscle Actin antibody - Smooth Muscle Cell Marker (ab5694)

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 - alpha smooth muscle Actin antibody - Smooth Muscle Cell Marker (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
Donkey anti rabbit (HRP) at 1/2500 dilution

Performed under reducing conditions.

Predicted band size : 42 kDa


This image is and 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.

Immunocytochemistry/ Immunofluorescence - alpha smooth muscle Actin antibody (ab5694)

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.

This image is courtesy of an Abreview submitted by Dr Jianyuan Chai

Immunocytochemistry/ Immunofluorescence - alpha smooth muscle Actin antibody (ab5694)

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.

This image is courtesy of an anonymous Abreview

Western blot - alpha smooth muscle Actin antibody (ab5694)

All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1/1000 dilution

Lane 1 : Lystates prepared from pig hear tissue from normal control animals
Lane 2 : Lystates prepared from pig hear tissue from normal control animals
Lane 3 : Lystates prepared from pig hear tissue from experimental animals
Lane 4 : Lystates prepared from pig hear tissue from experimental animals

Lysates/proteins at 4 µg per lane.

Secondary
HRP-conjugated goat polyclonal to rabbit IgG at 1/20000 dilution

Performed under reducing conditions.

Predicted band size : 42 kDa
Observed band size : 45 kDa (why is the actual band size different from the predicted?)


Exposure time : 1 minute

This image is a courtesy of Mario Torrado

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - alpha smooth muscle Actin antibody (ab5694)

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.

This image is courtesy of an Anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)

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.

Image courtesy of an anonymous Abreview.