Anti-TIAM1 antibody (ab54458)

  Western blot - TIAM1 antibody (ab54458)

All lanes : Anti-TIAM1 antibody (ab54458) at 1/1000 dilution

Lane 1 : NIH3T3 whole cell lysate at 50 µg
Lane 2 : Jurkat whole cell lysate at 25 µg


Predicted band size : 191 kDa
Observed band size : 191 kDa

  Immunocytochemistry/ Immunofluorescence - TIAM1 antibody (ab54458)

ICC/IF image of ab54458 stained HeLa 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 (ab54458, 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) - TIAM1 antibody (ab54458)

IHC image of ab54458 staining in human normal tonsil 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 ab54458, 10µg/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.