Anti-N Cadherin antibody - Intercellular Junction Marker

• WB

Ap

Western blot - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab18203 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

The N Cadherin protein has a predicted molecular weight of 100 kDa, however it is extensively glycosylated and has been shown to run in the 125-135 kDa region (SwissProt data).

All lanes:

Western blot - Anti-N Cadherin antibody - Intercellular Junction Marker (ab18203) at 1 µg/mL

Lane 1:

Brain (Rat) Tissue Lysate at 10 µg

Lane 2:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 3:

Brain (Human) Tissue Lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

Predicted band size: 99 kDa

Observed band size: 125 kDa

true

Exposure time: 1min

• IHC-P

AbReview34951****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

ab18203 staining N Cadherin in Human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a 10 mM citrate buffer pH6.0. Samples were incubated with primary antibody (1/100 in PBS plus casein) for 90 minutes at 37°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

Anti-N Cadherin antibody - Intercellular Junction Marker (ab18203) stained human embryonic stem cells differentiated into mesoderm.

• ICC/IF

AbReview60109****

Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

Paraformaldehyde-fixed, 0.2% Triton X100 permeabilized HaCaT (human keratinocyte cell line) cells stained for N Cadherin (green) using ab18203 at 1/200 dilution in ICC/IF, followed by Donkey anti Rabbit Alexa Fluor 568 at 1/500 dilution.

This image is courtesy of an Abreview submitted by Dr. Ann Wheeler.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

Immunohistochemical analysis of formalin fixed paraffin embedded human heart labelling N Cadherin with ab18203 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab18203 anti-N Cadherin antibody was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

Immunohistochemical analysis of formalin fixed paraffin embedded human testis labelling N Cadherin with ab18203 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab18203 anti-N Cadherin antibody was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

IHC image of N Cadherin staining in Human liver cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18203, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

Anti N-cadherin (ab18203) staining of human ovarian cancer tissue using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/1000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

Image courtesy of Mr Carl Hobbs, Kings College London.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

Anti N-cadherin (ab18203) staining of E17 developing rat retina using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/1000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

Image courtesy of Mr Carl Hobbs, Kings College London.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

Anti N-cadherin (ab18203) staining of mouse brain using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/1000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

Image courtesy of Mr Carl Hobbs, Kings College London.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

Anti N-cadherin (ab18203) staining in a human melanoma xenograft mouse model using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/1000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

Image courtesy of Mr Carl Hobbs, Kings College London.

• IP

Unknown

Immunoprecipitation - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

N Cadherin was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to N Cadherin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab18203.
Secondary : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band : 135kDa : N Cadherin

All lanes:

Immunoprecipitation - Anti-N Cadherin antibody - Intercellular Junction Marker (ab18203)

Predicted band size: 99 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

Immunohistochemistry of kidney carcinoma staining N Cadherin with ab18203 at 1μg/ml.