Anti-N Cadherin antibody [EPR19654] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [EPR19654] - BSA and Azide free (AB236032)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207608).

Heat mediated antigen retrieval was performed 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] - BSA and Azide free (AB236032)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207608).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-N Cadherin antibody [EPR19654] - BSA and Azide free (AB236032)

This data was developed using the same antibody clone in a different buffer formulation (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] (<a href='/en-us/products/primary-antibodies/n-cadherin-antibody-epr19654-ab207608'>ab207608</a>) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-N Cadherin antibody [EPR19654] - BSA and Azide free (ab236032) 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] - BSA and Azide free (AB236032)

This data was developed using ab207608, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/n-cadherin-antibody-epr19654-ab207608'>ab207608</a>) 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