HRP Anti-Sarcomeric Alpha Actinin antibody [EP2529Y]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (AB314407)

IHC image of Sarcomeric Alpha Actinin staining in a section of formalin-fixed paraffin-embedded normal human heart*. Performed on a Leica BOND™ system. The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. The section was then incubated with ab314407 at 10 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• WB

Lab

Western blot - HRP Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (AB314407)

Western blot : HRP Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab314407) staining at 1/5000 dilution, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab314407 was shown to bind specifically to ACTN2. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibody Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 2 minutes 30 seconds exposure time.

All lanes:

Western blot - HRP Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab314407) at 1/5000 dilution

Lane 1:

Human skeletal muscle lysate at 20 µg

Lane 2:

Mouse heart lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 103 kDa

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