Anti-E2F3 antibody (ab50917)

  Western blot - E2F3 antibody (ab50917)

Anti-E2F3 antibody (ab50917) at 0.25 µg/ml + Fetal thymus lysate at 10 µg

Secondary
HRP conjugated anti-Rabbit IgG at 1/50000 dilution

Predicted band size : 49 kDa
Observed band size : 39 kDa (why is the actual band size different from the predicted?)


Gel concentration: 12%

  Immunocytochemistry/ Immunofluorescence - E2F3 antibody (ab50917)

ICC/IF image of ab50917 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab50917, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-E2F3 antibody(ab50917)

IHC image of ab50917 staining in human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab50917, 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.