Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Timeless with ab72458 at 1/400 (0.5µg/ml). A DyLight® 594-conjugated goat anti-rabbit IgG (H+L) (1/100) was used as the secondary antibody.
All lanes : Anti-Timeless antibody (ab72458) at 0.04 µg/mlLane 1 : HeLa whole cell lysate at 50 µgLane 2 : HeLa whole cell lysate at 15 µgLane 3 : HeLa whole cell lysate at 5 µgLane 4 : 293T whole cell lysate at 50 µg
HeLa whole cell lysate, (1 mg for IP, 20% of IP loaded) with ab 72458 used for IP at 3 µg/mg lysate, and for subsequent WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 3 seconds.
ab72458 (1/200) staining Timeless in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100 and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.See Abreview
IHC image of ab72458 staining Timeless in Human Tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab72458, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.