Anti-CTNNA3 抗体 [EPR18307] (ab184916)

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling CTNNA3 with ab184916 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic and membrane staining on tumor cells of Human colonic adenocarcinoma is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cardiac muscle labelling Cardiac Troponin T with ab307152 at 1/5000 dilution (0.102 µg/mL) (B), CTNNA3 with ab184916 at 1/2000 dilution (0.942 μg/ml) (C) and PDystrophin with ab275391 at 1/500 dilution (2.08 μg/ml) (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A: merged staining of anti-Dystrophin (red; Opal™690), anti-Cardiac troponin T (green; Opal™520) and anti-CTNNA3 (gray; Opal™570) on human cardiac muscle.
Panel B: anti-Cardiac troponin T stained on cytoplasm of cardiac muscle.
Panel C: anti-CTNNA3 stained on intercalated discs.
Panel D: anti-Dystrophin stained on membrane of cardiac muscle.

The section was incubated in three rounds of staining: in the order of ab275391, ab307152, and ab184916 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling CTNNA3 with ab184916 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Intercalated disks staining on Mouse cardiac muscle is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling CTNNA3 with ab184916 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Intercalated disks staining of normal Human cardiac muscle is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling CTNNA3 with ab184916 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Intercalated disks staining on Rat cardiac muscle is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Blocking/Dilution buffer: 5% NFDM/TBST.

CTNNA3 was immunoprecipitated from 1mg of HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate with ab184916 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab184916 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1: HEK-293 whole cell lysate, 10 µg (Input).

Lane 2: ab184916 IP in HEK-293 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184916 in HEK-293 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 1 second