Western blot - N Cadherin antibody (ab18203)

All lanes : Anti-N Cadherin antibody (ab18203) at 1 µg/ml

Lane 1 : Rat Brain Lysate
Lane 2 : Mouse Brain Lysate
Lane 3 : Rat Brain Lysate with N Cadherin peptide (ab18620) at 1 µg/ml
Lane 4 : Mouse Brain Lysate with N Cadherin peptide (ab18620) at 1 µg/ml

Lysates/proteins at 20 µg per lane.

Secondary
Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 135 kDa
Observed band size : 135 kDa
Additional bands at : 50 kDa (possible truncated form).

Immunocytochemistry/ Immunofluorescence - N Cadherin antibody (ab18203)

anti-N cadherin antibody ab18203 stained human embryonic stem cells differentiated into mesoderm.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - N Cadherin antibody (ab18203)

IHC image of N Cadherin staining in human liver carcinoma FFPE section, performed on a Bond^TM 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.

Flow Cytometry - N Cadherin antibody (ab18203)

ab18203 staining CD63 in human Fibrosarcoma HT1080 cell line by flow cytometry. Cells were incubated with primary antibody for 1 hour. ab7007, a donkey polyclonal to rabbit Ig, was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

Immunoprecipitation - Anti-N Cadherin antibody (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; non specific - 70 and 50kDa: We are unsure as to the identity of this extra band.