Cultured human endothelial cells were labeled with [6-3H]gluco-samine and subsequently extracted with 1% Triton X-100 in buffer as described in Material and methods
The use of 1% Triton X-100 causes a release of about 70–80% of the cadherin/ catenin complex (Schnittler ) that can be immunoprecipitated from a 8000 × g supernatant using a monoclonal VE-cadherin antibody
Precipitated protein was subjected to SDS-PAGE, and fluorography of the gel displayed one main band at MW of 135 kDa and few weaker labeled bands (Figure 1)
Part of the precipitate was subjected to Western blot analysis using a pan cadherin antibody known to react with a short carboxyterminal sequence of classical cadherins
This antibody stained a single band exactly at the same MW as the main radiolabeled band (data not shown)