Figure 2
From: Application of Gaussia luciferase in bicistronic and non-conventional secretion reporter constructs
Bicistronic expression system for release of Gaussia luciferase activity. A, whole cell lysates from transfected 293 T cells described in Figure 1E were subjected to 10% PAGE and subsequent immunoblotting using a rabbit anti-GLUC antibody. Bands corresponding to GLUC are indicated by an arrowhead and bands corresponding to intact GFP-SPGLUC and β-actin-SPGLUC are indicated by asterisks. B, 293 T cells transfected with GLUC or GFP-SPGLUC were analyzed for GLUC activity in cell lysates and supernatants by immuno-blotting with anti-GLUC antibody. Supernatants and cell lysates were collected from the same well. Supernatants were enriched as described in Methods. Cell lysates and supernatants were resolved by PAGE and bands were detected with anti-GLUC antibody. Bands corresponding to GLUC are indicated by an arrowhead and bands corresponding to intact GFP-SPGLUC are indicated by an asterisk. Experiments have been performed three times and shown are representative blots. SN – supernatant.