Anti-N Cadherin antibody [EPR19654]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR19654] (AB207608)

Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue labeling N Cadherin with ab207608 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Intercalated disc staining on human cardiac muscle is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR19654] (AB207608)

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling N Cadherin with ab207608 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on human endometrium is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-N Cadherin antibody [EPR19654] (AB207608)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1/2 : 30 seconds; Lane 3 : 5 seconds; Lane 4/5 : 3 minutes.

The molecular weight observed is consistent with what has been described in the literature(PMID : 22553038).

All lanes:

Western blot - Anti-N Cadherin antibody [EPR19654] (ab207608) at 1/1000 dilution

Lane 1:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 10 µg

Lane 2:

PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 3:

Human fetal brain lysate at 10 µg

Lane 4:

Human cerebellum lysate at 10 µg

Lane 5:

Human fetal liver lysate at 10 µg

Secondary

Lanes 1 - 2:

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 1/100000 dilution

Lanes 3 - 5:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 99 kDa

Observed band size: 110 kDa,130 kDa

false

• WB

Lab

Western blot - Anti-N Cadherin antibody [EPR19654] (AB207608)

False colour image of Western blot : Anti-N Cadherin antibody [EPR19654] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab207608 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 [EPR19654] (ab207608) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-N Cadherin antibody [EPR19654] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/n-cadherin-antibody-epr19654-bsa-and-azide-free-ab236032'>ab236032</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

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

Lane 2:

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

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 [EPR19654] (AB207608)

Exposure time : Lane 1 and 2 : 4 seconds, Lane 4 to 11 : 1 seconds.

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)

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

All lanes:

Western blot - Anti-N Cadherin antibody [EPR19654] (ab207608) 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 [EPR19654] (AB207608)

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

ab207608 shows weak cross reactivity with R Cadherin.

Lanes 1 - 2:

Western blot - Anti-N Cadherin antibody [EPR19654] (ab207608) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (<a href='/en-us/products/primary-antibodies/6x-his-tag-antibody-epr20547-chip-grade-ab213204'>ab213204</a>)

Lanes 1 and 3:

His-tagged Human N Cadherin recombinant protein (aa160-724) at 10 ng

Lanes 2 and 4:

His-tagged Human R Cadherin recombinant protein (aa170-734) at 10 EU/ng

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

Observed band size: 75-100 kDa

false

Exposure time: 80s