Detection of the G-CSFR protein in Western blot analyses. In each case, 2 × 107 C243 cells stably transfected with the human wild-type G-CSFR or different deletion mutants (D1-D3) were lysed. The G-CSF receptors were immunoprecipitated with MoAb 129, separated by SDS-PAGE using an 8% wt/vol polyacrylamide gel, and blotted onto nitrocellulose membranes. Detection was performed with anti–G-CSFR antibody (MoAb 129; IgG1) and horseradish peroxidase (HRP)-conjugated goat-antimouse IgG. The immunoblot was developed by the enhanced chemiluminescence method following the manufacturer's guidelines (ECL; Amersham, Braunschweig, Germany). Lane 1, wild-type G-CSFR (aa 1-813); lane 2, deletion mutant D1 (aa 1-670); lane 3, deletion mutant D2 (aa 1-685); lane 4, deletion mutant D3 (aa 1-634).
