Anti-N Cadherin antibody [EPR1791-4]

• WB

Lab

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] (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 (<a href='/en-us/products/primary-antibodies/n-cadherin-antibody-epr1791-4-bsa-and-azide-free-ab271856'>ab271856</a>)

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

• IHC-P

Lab

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 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.

• IHC-P

Lab

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 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.

• IHC-P

Lab

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 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.

This data was developed using the same antibody clone in a different buffer formulation (ab271856).

• WB

Lab

Western blot - Anti-N Cadherin antibody [EPR1791-4] (AB76011)

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] (ab76011) 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

Lab

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, 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 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] (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

Lane 2:

Western blot - Human CDH2 (N Cadherin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-cdh2-n-cadherin-knockout-hek-293t-cell-line-ab255377'>ab255377</a>)

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 99 kDa

Observed band size: 100 kDa,125 kDa

false

• WB

Lab

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 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] (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

Predicted band size: 99 kDa

Observed band size: 125 kDa

false

• IHC-P

Unknown

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.

• WB

CiteAb

Western blot - Anti-N Cadherin antibody [EPR1791-4] (AB76011)

Western Blotting using Anti-N Cadherin antibody [EPR1791-4], ab76011. Publication image from Liu, S. C. et al., 2018, Nat Commun, 30504771. Legend direct from paper.

AZD0530 treatment suppresses LIF-mediated tumor invasion. a Western blot analysis of YAP1 and focal adhesion proteins in cancer cells treated with AZD0530 (5 µM). Protein lysates were harvested at 24 h post treatment. b Immunostaining for YAP1 (red) in cancer cells treated with AZD0530. Alexa Fluor 488 phalloidin (green) was used to stain F-actin. Blue, nuclear staining. Scale bars, 10 µm. c–e Representative images of YAP1 (c), p-PXN (Y118) (d), and p-FAK (Y397) (e) expression in mouse WT and cLIF xenografts treated with AZD0530 or vehicle. Scale bars, 50 µm. f Quantification of mouse NPC xenografts with events of local invasion based on results of hematoxylin and eosin staining. The AZD0530 treatment procedure in the mouse model is described in Methods

false

• WB

CiteAb

Western blot - Anti-N Cadherin antibody [EPR1791-4] (AB76011)

Western Blotting using Anti-N Cadherin antibody [EPR1791-4], ab76011. Publication image from Liu, S. C. et al., 2018, Nat Commun, 30504771. Legend direct from paper.

Cytoplasmic LIF regulates EMT and invadopodia formation. a Migration pattern analysis using time-lapse live imaging in wound-healing experiments. Scale bars, 100 µm. See also Supplementary movies 1–3. b 3D-gelatin invasion assays. The invasion depth of cancer cells through a stiff gelatin matrix (6%) were measured by time-lapse phase contrast vertical scanning (Olympus IX83). Values are presented as means ± SD of triplicate experiments. c Representative IHC images of LIF expression in paraffin-embedded mouse tumor xenografts. Scale bars, 50 µm. d Quantification of mouse tumor xenografts with events of local invasion based on hematoxylin & eosin (HE) staining analysis. e Western blot of EMT and invadopodia markers in wild-type parental, cLIF and LIF+/− cancer cells. GAPDH was used as a loading control. f–h Immunofluorescence staining of invadopodia markers : TKS5 (f). CTTN (g). MMP2 (h). Alexa Fluor 488 phalloidin (green) was used to stain F-actin. Blue, nuclear staining. Scale bars, 10 µm

false