Immunofluorescence microscopy of Saos-2 cells transduced with a retroviral vector expressing hTERT and green fluorescent protein (GFP) from an internal ribosomal entry site (IRES). Panel A shows native GFP expression (green), Panel B shows DAPI staining of chromsomes (blue), Panel C shows anti-hTERT staining at a 1:500 dilution followed by washes and addition of a 1:1000 dilution of rhodamine Goat anti-Rabbit IgG (code 611-1002) for detection. Panel D shows no staining of hTERT-transduced cells using pre-immune serum.

Immunofluorescence of Rockland anti-hTERT antibody - Two different lots of Rockland anti hTERT antibody were used to stain hTERT on hTERT-over-expressing fibroblasts. Cells were fixed in 4% PFA (in PBS) for 10 min and frozen in -80 after 3 min air-drying before incubation with Rockland anti hTERT 1:2000 overnight and staining with a 1:2000 dilution of Alexafluor488 secondary Ab. Confocal images provided by G. Saretzki, Institute for Ageing and Health, Newcastle University, UK. See Ahmed et. Al. for more information.

Immunofluorescence of Rockland anti-hTERT antibody was used to stain hTERT on hTERT-over-expressing fibroblasts. Cells were untreated (Left) or treated (Right) with 500 uM H202, fixed in 4% PFA (in PBS) for 10 min and frozen in -80 after 3 min air-drying before staining with Rockland anti-hTERT 1:2000 overnight. Confocal images provided by G. Saretzki, Institute for Ageing and Health, Newcastle University, UK. See Ahmed et. Al. for more information.

Western blot using anti-hTERT antibody (1:500 dilution).  Endogenous levels of mTERT in NIH 3T3 cells (lane 1) and hTERT in HeLa cells (lane 5) are not detectable.  After transduction with an hTERT expression vector, both cell types show high levels of hTERT protein (lanes 2 and 6).  SY5Y cells, which have high endogenous levels of hTERT, have detectable hTERT protein in both untransduced (lane 3) and transduced (lane 4) cells.  The arrow indicates a molecular weight of approximately 127kD, the expected size of hTERT protein.