Anti-N Cadherin antibody [EPR1791-4]

9 product images

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  This image is courtesy of an anonymous customer review.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR1791-4] (ab76011)

  ab76011 staining N Cadherin in Mouse pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA + 1% FBS for 2 hours at room temperature; antigen retrieval was by heat mediation in a citrate buffer pH6. Samples were incubated with primary antibody (1/500 in 1% BSA + 1% FBS) for 16 hours at 4°C. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

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  Western blot - Anti-N Cadherin antibody [EPR1791-4] (ab76011)

  This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab76011 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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-N Cadherin antibody [EPR1791-4] (ab76011) at 1/1000 dilution

  Lane 1: HeLa Whole Cell Lysate at 20 µg

  Lane 2: HeLa Whole Cell Lysate (Scraped) at 20 µg

  Lane 3: Human Brain Tissue Lysate at 20 µg

  Lane 4: Mouse Brain Tissue Lysate at 20 µg

  Lane 5: Rat Brain Tissue Lysate at 20 µg

  Lane 6: MCF7 Whole Cell Lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 99 kDa

  Observed band size: 125 kDa

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  Western blot - Anti-N Cadherin antibody [EPR1791-4] (ab76011)

  Lanes 1-2: Merged signal (red and green). Green - ab76011 observed at 100 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
  

  ab76011 was shown to react with N Cadherin in wild-type HEK-293T cells in Western blot. Loss of signal was observed when knockout sample Human CDH2 (N Cadherin) knockout HEK-293T cell lysate ab263843 was used. Wild-type and N Cadherin knockout samples were subjected to SDS-PAGE. ab76011 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 5000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-N Cadherin antibody [EPR1791-4] (ab76011) at 1/5000 dilution

  Lane 1: Wild-type HEK-293T cell lysate at 20 µg

  Lane 2: CDH2 knockout HEK-293T cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 99 kDa

  Observed band size: 100 kDa, 125 kDa

  Lanes 1 - 2: Merged signal (red and green). Green - ab76011 observed at 125 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
  

  ab76011 was shown to react with N Cadherin in wild-type HEK-293T. Loss of signal was observed when knockout cell line Human CDH2 (N Cadherin) knockout HEK-293T cell line ab255377 (knockout cell lysate Human CDH2 (N Cadherin) knockout HEK-293T cell lysate ab263843) was used. Wild-type and N Cadherin knockout samples were subjected to SDS-PAGE. ab76011 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 5000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

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  Western blot - Anti-N Cadherin antibody [EPR1791-4] (ab76011)

  The molecular weight observed is consistent with what has been described in the literature (PMID: 22553038). This antibody fails to detect N Cadherin in HCT 116 cell which is positive described in the literature (PMID: 23431386 and 26540342)

  All lanes: ab76011, Anti-N Cadherin antibody [EPR1791-4] (Left) or Anti-N Cadherin antibody [EPR19654] ab207608, Anti-N Cadherin antibody [EPR19654] (Right) at 1/1000 dilution

  Lane 1: A549 (Human lung carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method at 20 µg

  Lane 2: PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method at 20 µg

  Lane 3: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method at 20 µg

  Lane 4: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method at 20 µg

  Lane 5: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg

  Lane 6: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate prepared in 1%SDS Hot lysis method at 20 µg

  Lane 7: C6 (Rat glial tumor glial cell) whole cell lysates prepared in RIPA lysis method at 20 µg

  Lane 8: C6 (Rat glial tumor glial cell) whole cell lysates prepared in 1%SDS Hot lysis method at 20 µg

  Lane 9: Human brain lysates prepared in 1%SDS Hot lysis method at 20 µg

  Lane 10: Mouse brain lysates prepared in 1%SDS Hot lysis method at 20 µg

  Lane 11: Rat brain lysates prepared in 1%SDS Hot lysis method 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: 99 kDa

  Observed band size: 110-130 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR1791-4] (ab76011)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cardiac muscle tissue sections labeling N Cadherin with purified ab76011 at 1:50 dilution (1.94 μg/ml). Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR1791-4] (ab76011)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling N Cadherin with purified ab76011 at 1:50 dilution (1.94 μg/ml). Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR1791-4] (ab76011)

  Immunohistochemistry of kidney carcinoma staining N Cadherin with ab76011 at 1μg/ml
  

  Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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  Western blot - Anti-N Cadherin antibody [EPR1791-4] (ab76011)

  False colour image of Western blot: Anti-N Cadherin antibody [EPR1791-4] staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab76011 was shown to bind specifically to N Cadherin. A band was observed at 125 kDa in wild-type HeLa cell lysates with no signal observed at this size in cdh2 knockout cell line ab274934 (knockout cell lysate ab274992).
  
  To generate this image, wild-type and cdh2 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  Lanes 1 - 2: Western blot - Anti-N Cadherin antibody [EPR1791-4] (ab76011) at 1/5000 dilution

  Lanes 1 - 2: Western blot - Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free (Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free ab271856)

  Lanes 1 - 2: Western blot - Anti-N Cadherin antibody [EPR1791-4] - Low endotoxin, Azide free (Anti-N Cadherin antibody [EPR1791-4] - Low endotoxin, Azide free ab202030) at 1/5000 dilution

  Lane 1: Wild-type HeLa cell lysate at 20 µg

  Lane 2: cdh2 knockout HeLa cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 125 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR1791-4] (ab76011)

  Immunohistochemical analysis of formalin-fixed paraffin-embedded human heart labelling N Cadherin with Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free ab271856 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free ab271856 anti N Cadherin antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

  This data was developed using the same antibody clone in a different buffer formulation (Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free ab271856).