ab190685 at 1/100 dilution immunoprecipitating CTNNA1 in Jurkat HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+): HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab51032 in HeLa whole cell lysate
For western blotting, ab51032 at 1/500 dilution and VeriBlot for IP Detection Reagent (HRP)(ab131366) at 1/1000 dilution were used.

Blocking and diluting buffer: 5% NFDM /TBST.

Paraffin-embedded rat stomach tissue stained for CTNNA1 with ab51032 at a 1/100 dilution in immunohistochemical analysis. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody and Hematoxylin used as a counterstain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was performed for 20 minutes.

Positive staining was seen on rat stomach.

The section was incubated with ab51032 for 30 minutes at room temperature.

The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

Paraffin-embedded mouse liver tissue stained for CTNNA1 with ab51032 at a 1/100 dilution in immunohistochemical analysis. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody and Hematoxylin used as a counterstain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was performed for 20 minutes.

Positive staining was seen on mouse liver.

The section was incubated with ab51032 for 30 minutes at room temperature.

The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

Paraffin-embedded human stomach tissue stained for CTNNA1 with ab51032 at a 1/100 dilution in immunohistochemical analysis. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody and Hematoxylin used as a counterstain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was performed for 20 minutes.

Positive staining was seen on human stomach.

The section was incubated with ab51032 for 30 minutes at room temperature.

The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

All lanes : Anti-CTNNA1 antibody [EP1793Y] (ab51032) at 1/10000 dilution

Lane 1 : Mouse heart lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat kidney lysate
Lane 5 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 6 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 7 : HUVEC (Human umbilical vein endothelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 100 kDa
Observed band size: 100 kDa

Expsure time

Lane 1-4: 180 seconds

Lane 5,7: 40 seconds

Lane 6: 5 seconds

Blocking/diluting buffer and concentration: 5% NFDM/TBST

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: CTNNA1 HAP1 whole cell lysate (20 µg) 
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab51032 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab51032 was shown to recognize CTNNA1 in wild-type cells as signal was lost at the expected MW in CTNNA1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CTNNA1 knockout samples were subjected to SDS-PAGE. Ab51032 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/50000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.