Flow cytometry overlay histogram showing Jurkat cells (human T cell leukemia T lymphocyte from peripheral blood) fixed with 4% paraformaldehyde and permeabilised with 90% methanol, stained with ab124964 at 1/1000 dilution (0.05 μg) red line. Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) secondary antibody used at a 1/5000 dilution.

Isotype control: Rabbit monoclonal IgG (ab172730) black line.

Unlabelled control: Cells without incubation with primary antibody and secondary antibody blue line.

Immunohistochemical analysis of formalin-fixed paraffin-embedded human endometrium labelling alpha smooth muscle actin with ab124964 at a concentration of 0.07µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab124964 anti alpha smooth muscle actin antibody was incubated at 37°C for 16min. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Tissue Microarrays stained for Anti-alpha smooth muscle Actin antibody [EPR5368] using ab124964 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested. The section was incubated with ab124964 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse stomach tissue labelling alpha smooth muscl Actin with ab124964 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

Positive staining on smooth muscle in mouse stomach.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labelling alpha smooth muscl Actin with purified ab124964 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.