Anti-CTNNA3 antibody [EPR18307]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTNNA3 antibody [EPR18307] (AB184916)

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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CTNNA3 antibody [EPR18307] (AB184916)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTNNA3 antibody [EPR18307] (AB184916)

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.

• IP

Supplier Data

Immunoprecipitation - Anti-CTNNA3 antibody [EPR18307] (AB184916)

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

All lanes:

Immunoprecipitation - Anti-CTNNA3 antibody [EPR18307] (ab184916)

Predicted band size: 100 kDa

false

Exposure time: 1s

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CTNNA3 antibody [EPR18307] (AB184916)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse cardiac muscle tissue staining Titin with ab307446 at a 1/1000 ( 0.475 µg/ml) dilution, CTNNA3 with ab184916 at 1/2000 ( 0.942 µg/ml) dilution and Dystrophin with ab218198 at 1/5000 ( 0.110 µg/ml) dilution.

Panel A : merged staining of anti-Titin (gray; Opal™570), anti-CTNNA3 (green; Opal™520) and anti-Dystrophin (magenta; Opal™690) on mouse cardiac muscle.
Panel B : anti-Titin staining nucleus and cytoplasm in mouse cardiac muscle.
Panel C : anti-CTNNA3 staining intercalated discs in mouse cardiac muscle.
Panel D : anti-Dystrophin staining membrane in mouse cardiac muscle.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab307446, ab184916 and ab218198 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.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTNNA3 antibody [EPR18307] (AB184916)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTNNA3 antibody [EPR18307] (AB184916)

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.

• WB

Supplier Data

Western blot - Anti-CTNNA3 antibody [EPR18307] (AB184916)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CTNNA3 antibody [EPR18307] (ab184916) at 1/5000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 100 kDa

Observed band size: 100 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-CTNNA3 antibody [EPR18307] (AB184916)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CTNNA3 antibody [EPR18307] (ab184916) at 1/50000 dilution

Lane 1:

HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

C2C12 (Mouse myoblast cell line) whole cell lysate at 20 µg

Lane 3:

Rat1 (Rat fibroblast cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 100 kDa

Observed band size: 100 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-CTNNA3 antibody [EPR18307] (AB184916)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CTNNA3 antibody [EPR18307] (ab184916) at 1/5000 dilution

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

Mouse heart tissue lysate at 10 µg

Lane 3:

Rat brain tissue lysate at 10 µg

Lane 4:

Rat heart tissue lysate at 10 µg

Lane 5:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 6:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 7:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 100 kDa

Observed band size: 100 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-CTNNA3 antibody [EPR18307] (AB184916)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CTNNA3 antibody [EPR18307] (ab184916) at 1/50000 dilution

All lanes:

MDCK (Canine kidney cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 100 kDa

Observed band size: 100 kDa

false

Exposure time: 1s