Western and ligand blots of the recombinant VHDL receptor

Copyright information:Taken from "Cloning and expression of the VHDL receptor from fat body of the corn ear worm, "Journal of Insect Science 2004;4():-.Published online 27 Feb 2004PMCID:PMC455680.Copyright © 2004. Open access; copyright is maintained by the authors. Sf9 cells were transfected with a VHDL-receptor cDNA expression vector, as described in Materials and Methods. Transfected and wild-type cells (control) were lysed, and the presence of VHDL receptor analysed in the growth medium and the cell lysate. Panel A: Dot blots of 3 μg of cell lysate (lysate) or concentrated growth medium (medium) were spotted onto PVDF membrane, and processed as described in Materials and Methods. The Western blot was probed with anti-VHDL-receptor antiserum (1:6,000, secondary antibody 1:10,000; Western). Ligand blots were carried out with biotinylated VHDL (1: 8,000, Ligand Biotin-VHDL) or FITC-labeled VHDL (1:8000, Ligand FITC-VHDL) Panel B: Ligand blot with FITC-VDHL following electrophoresis. Left lane: purified receptor (0.1 μg), right lane: protein concentrated from the Sf9-lysate (3 μg).