This is a knockout-validated antibody summary, based on the publication "N-cadherin regulates signaling mechanisms required for lens fiber cell elongation and lens morphogenesis", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG1

Company: BD Biosciences

Antibody: N-Cadherin

Catalog number: 610921

Summary: Mouse monoclonal IgG1 against mouse N-Cadherin (aa. 802-819). Reacts with human, mouse, rat, and chicken. Suitable for western blot, immunofluorescence and immunoprecipitation.

Western blot | Immunohistochemistry

WB: E16.5 wildtype and N-cadcKO lenses isolated from embryos. Samples were lysed in Triton X-100/Octylglucoside (OG/T) buffer cocktail composed of 44.4 mM n-octyl β-D-glucopyranoside, 1% Triton X-100, 100 mM NaCl, 1 mM MgCl2, 5 mM EDTA, and 10 mM imidazole, containing 1 mM sodium vanadate, 0.2 mM H2O2, and protease inhibitor cocktail (Sigma Aldrich, P8340).

IHC: Isolated mouse eyes were fixed in 3.7% formaldehyde overnight at 4 °C, cryoprotected in 30% sucrose solution for a minimum of 24 h prior to freezing and 20-μm thick cryosections cut.

WB: 5% skim milk for 1 h.

IHC: 5% goat serum, 0.5 g BSA in 50 ml DPBS for 1 h

WB: at 4 °C overnight.

IHC: at 4 °C overnight.

WB: secondary antibodies conjugated to horseradish peroxidase (BIO-RAD, 170-6515, 170-6516).

IHC: fluorescent-conjugated secondary antibody for 1 h at 37°C (Jackson ImmunoResearch Laboratories, 111-295-144, 115-545-003, 115-295-008).

WB: ECL reagent or ECL plus reagent (Thermo Fisher Scientific Inc. Rockford, 32106, 80197) and images were acquired using the FluorChem E & M imager from Protein Simple (FM0418).

IHC: Confocal microscopy was performed using a Zeiss LSM510META confocal microscope.

Cryosections of E13.5 wildtype (A) and N-cadcKO (B) eyes labeled for N-cadherin show loss of N-cadherin in N-cadcKO lenses by E13.5. (C) Immunoblot for N-cadherin in E16.5 wildtype and N-cadcKO lenses confirms the loss of N-cadherin protein in the N-cadcKO lens, with GAPDH as a loading control. Please see Figure 2a-c in the article [1].