Immunocytochemistry/ Immunofluorescence - alpha smooth muscle Actin antibody [1A4] (ab7817)

ab7817 staining alpha smooth muscle Actin in mouse heart cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with TritonX-100 and blocked with 5% BSA for 30 minutes at room temperature. Samples were incubated with primary antibody 1/100 in blocking buffer for 2 hours. An Alexa Fluor^® 488-conjugated Donkey monoclonal to mouse IgG, dilution 1/200, was used as secondary antibody.

This image is courtesy of an Abreview submitted by Dr. Ho-Jae Lee

Immunocytochemistry/ Immunofluorescence - alpha smooth muscle Actin antibody [1A4] (ab7817)

Cannabinoid receptor (CB1) (green) and α-smooth muscle actin immunoreactivity (red) in a cross-section of rat middle cerebral artery. Rat middle cerebral artery sections were incubated for 12 hours with a 1:300 dilution of anti-CB1 polyclonal antibody raised in rabbits and a 1:100 dilution of anti-α-smooth muscle actin monoclonal antibody raised in mice. The secondary antibodies were Alexa-488-labeled goat anti-rabbit at a 1:500 dilution and Cy3-labeled goat anti-mouse at a 1:1,000 dilution. This picture was submitted by David Rademacher whose review of the antibody is in the review section of this datasheet.

Western blot - alpha smooth muscle Actin antibody [1A4] (ab7817)

All lanes : Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1/200 dilution

Lane 1 : HeLa Nuclear
Lane 2 : HeLa whole cell lysate
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-mouse at 1/5000 dilution

Performed under reducing conditions.

Observed band size : 40 kDa (why is the actual band size different from the predicted?)
Additional bands at : 37 kDa. We are unsure as to the identity of these extra bands.

Fluorescence detection of secondary antibody.

Immunocytochemistry - alpha smooth muscle Actin antibody [1A4] (ab7817)

Ab7817 positively staining paraformaldehyde fixed porcine valvular interstitial cells (VICs) at 1/75. Staining was carried out in conjunction with goat anti mouse Alexa Ò488 (1/400).

Ab 7817 was used to detect myofibroblsats among a heterogeneous interstitial cell type. The image shows heparin treated VICs labelled for alpha actin (green), and counterstained with propidium iodide (red).

This image is an edited version of an image received courtesy of an Abreview on on 22 August 2005. We do not have any further information relating to this image.