Human/Mouse/Rat N‑Cadherin Antibody

Human N‑Cadherin ELISA Standard Curve.

Recombinant Human N-Cadherin protein was serially diluted 2-fold and captured by Mouse Anti-Human N-Cadherin Monoclonal Antibody (Catalog # MAB13883) coated on a Clear Polystyrene Microplate (Catalog # DY990). Sheep Anti-Human/Mouse/Rat N-Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6426) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Detection of N‑Cadherin in A172 cells by Flow Cytometry.

A172 cells were stained with Sheep Anti-Human/Mouse/Rat N‑Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6426, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Fluorescein-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0125). View our protocol for Staining Membrane-associated Proteins.

Detection of Human N-Cadherin by Western Blot

N-Cadherin Depletion Rescues the Migration Defect in Rab14- and FAM116-Depleted Cells(A and B) A549 cells were treated with control, Rab14, FAM116A, N-cadherin, E-cadherin, and FGFR2 siRNA duplexes alone or in the combinations indicated for 72 hr (A). The cells were then western blotted for N-cadherin, E-cadherin, or tubulin as a loading control or (B) fixed and then stained with DAPI and antibodies to N-cadherin. Scale bar indicates 10 μm.(C) A549 cells were treated with control, Rab14, FAM116A, N-cadherin, E-cadherin, and FGFR2 siRNA duplexes alone or in the combinations indicated for 72 hr. The cell monolayers were then scratched and imaged for 16 hr. Cell migration was measured and is plotted on the graph with error bars to show the standard error of the mean (n = 3).(D) Images are shown from the 0 and 16 hr time points for the Rab14- and FAM116A-depleted cells in the presence and absence of N-cadherin siRNA. Scale bar indicates 50 μm.See also Figure S5. Image collected and cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S1534580712001876), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human N-Cadherin by Western Blot

Abnormal Adherens Junctions in Rab14- and FAM116-Depleted Cells(A) A549 cells were treated with control, Rab4, Rab11, Rab14, FAM116A, and Avl9 siRNA duplexes for 72 hr. The cells were lysed in SDS-PAGE sample buffer and then western blotted as indicated in the figure. Tubulin was used as a loading control.(B) A549 cells were treated with control, Rab14, and FAM116A siRNA duplexes for 72 hr, fixed, and then stained with DAPI and antibodies to N-cadherin.(C) A549 cells were treated with control, Rab4, Rab11, Rab14, FAM116A, and Avl9 siRNA duplexes for 72 hr. The cell monolayers were then scratched, samples fixed at 0, 4, 8, and 16 hr, and then stained with DAPI and antibodies to N-cadherin. Images are oriented so the wound is to the right. Scale bar indicates 10 μm in all images.See also Figure S6. Image collected and cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S1534580712001876), licensed under a CC-BY license. Not internally tested by R&D Systems.