Anti-N Cadherin antibody - Intercellular Junction Marker

11 product images

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  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 ECL Substrate Kit (High Sensitivity) 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

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 99 kDa

  Observed band size: 125 kDa

  Exposure time: 1min

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  This image is courtesy of an anonymous customer review

  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.

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  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) (Mouse monoclonal [SB62a] Anti-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

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  Image courtesy of Mr Carl Hobbs, Kings College London.

  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.

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  Image courtesy of Mr Carl Hobbs, Kings College London.

  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.

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  Image courtesy of Mr Carl Hobbs, Kings College London.

  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.

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  Image courtesy of Mr Carl Hobbs, Kings College London.

  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.

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  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.

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  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.

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  This image is courtesy of a customer review submitted by Dr. Ann Wheeler.

  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.

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  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.