Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free (AB271856)

Immunohistochemical analysis of formalin-fixed paraffin-embedded human heart labelling N Cadherin with 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). 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free (AB271856)

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 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76011).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free (AB271856)

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 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76011).

• WB

Unknown

Western blot - Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free (AB271856)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 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 (ab216773) and 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] - BSA and Azide free (ab271856) 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

Predicted band size: 99 kDa

Observed band size: 125 kDa

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• WB

Lab

Western blot - Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free (AB271856)

This data was developed using the same antibody clone in a different buffer formulation (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] (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] (<a href='/en-us/products/primary-antibodies/n-cadherin-antibody-epr1791-4-ab76011'>ab76011</a>) at 1/5000 dilution

Lanes 1 - 2:

Western blot - 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 (<a href='/en-us/products/primary-antibodies/n-cadherin-antibody-epr1791-4-low-endotoxin-azide-free-ab202030'>ab202030</a>) at 1/5000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CDH2 (N Cadherin) knockout HeLa cell lysate (ab274992)

Lane 2:

Western blot - Human CDH2 (N cadherin) knockout HeLa cell line (ab274934)

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

Observed band size: 125 kDa

false

• WB

Lab

Western blot - Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free (AB271856)

This data was developed using ab76011, the same antibody clone in a different buffer formulation.

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

Lanes 1 - 2 : Merged signal (red and green). Green - ab76011 observed at 125 kDa. Red - 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 ab255377 (knockout 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 (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 (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/n-cadherin-antibody-epr1791-4-ab76011'>ab76011</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 99 kDa

Observed band size: 110-130 kDa

false

• WB

Unknown

Western blot - Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free (AB271856)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76011).

Lanes 1 - 2 : Merged signal (red and green). Green - ab76011 observed at 125 kDa. Red - 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 ab255377 (knockout 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 (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 (ab216773) and 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] - BSA and Azide free (ab271856) 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

Predicted band size: 99 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR1791-4] - BSA and Azide free (AB271856)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76011).