Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)

Transgenic mice expressing GFP selectively in lamina II of the spinal cord.

In the right panels, note the correspondance between the green (rabbit anti-GFP) and red signals (chicken anti-GFP from Abcam) indicating that these two antibody preparations recognized the same gene product. The secondary antibody used with ab13970 was an FITC-labeled goat anti-chicken

Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)

ab13970 staining GFP in GFP-transfected NIH/3T3 (Mouse embryo fibroblast cell line) cells.

The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab13970 at 1/2000 dilution overnight at +4°C followed by incubation with Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173), for 1 hour, at 1 μg/ml.

Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH/3T3 cells, the cells upon which ab13970 was applied gave a stronger signal in the 488 channel, indicating that ab13970 is binding to GFP and therefore eliciting signal amplification.

ab13970 was also applied to non-GFP-transfected NIH/3T3 cells, which produced no positive staining, indicating specificity for GFP.

Nuclear DNA was labeled with 1.43 μM DAPI (blue).