Anti-CTNNA1 antibody [EP1793Y]

7 product images

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  Immunoprecipitation - Anti-CTNNA1 antibody [EP1793Y] (ab51032)

  Anti-GCLC antibody [EP13475] 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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab51032 in HeLa whole cell lysate
  For western blotting, ab51032 at 1/500 dilution and VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution were used.
  

  Blocking and diluting buffer: 5% NFDM /TBST.

  All lanes: Immunoprecipitation - Anti-CTNNA1 antibody [EP1793Y] (ab51032)

  Predicted band size: 100 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTNNA1 antibody [EP1793Y] (ab51032)

  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 (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.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTNNA1 antibody [EP1793Y] (ab51032)

  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 (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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTNNA1 antibody [EP1793Y] (ab51032)

  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 (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.

• 

  Western blot - Anti-CTNNA1 antibody [EP1793Y] (ab51032)

  Expsure time
  

  Lane 1-4: 180 seconds

  Lane 5,7: 40 seconds

  Lane 6: 5 seconds

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

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

  Lane 1: Mouse heart lysate at 20 µg

  Lane 2: Mouse kidney lysate at 20 µg

  Lane 3: Rat brain lysate at 20 µg

  Lane 4: Rat kidney lysate at 20 µg

  Lane 5: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 6: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 7: HUVEC (Human umbilical vein endothelial cell) whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 100 kDa

  Observed band size: 100 kDa

• 

  Western blot - Anti-CTNNA1 antibody [EP1793Y] (ab51032)

  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, Anti-GAPDH antibody [mAbcam 9484] - 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 Anti-GAPDH antibody [mAbcam 9484] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-CTNNA1 antibody [EP1793Y] (ab51032)

  Predicted band size: 100 kDa

• 

  Western blot - Anti-CTNNA1 antibody [EP1793Y] (ab51032)

  All lanes: Western blot - Anti-CTNNA1 antibody [EP1793Y] (ab51032) at 1/50000 dilution

  All lanes: Western blot - Recombinant Human CTNNA1 protein (Tagged) (Recombinant Human CTNNA1 protein (Tagged) ab51443) at 0.01 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ab97080) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 100 kDa

  Exposure time: 1min