Anti-STAU2 antibody (ab60724)

Western blot - STAU2 antibody (ab60724)

Anti-STAU2 antibody (ab60724) at 1 µg/ml + IMR-32 cell lysate at 25 µg

Predicted band size : 63 kDa
Observed band size : 53 kDa (why is the actual band size different from the predicted?)
Additional bands at : 35 kDa. We are unsure as to the identity of these extra bands.

Immunocytochemistry/ Immunofluorescence - STAU2 antibody (ab60724)

Cell-derived microvesicles (from human bone marrow derived mesenchymal stem cells and liver resident stem cells) were fixed in 4% paraformaldheyde in PBS containing 2% sucrose for 15 minutes and permeabilized with cold methanol (-20ºC). After blocking with 1% BSA in PBS, samples were incubated with ab60724 at a 1/100 dilution. After washings, cells were incubated with the appropriate secondary antibodies at 1/1000 dilution.

Image from Collino F et al, PLoS One. 2010 Jul 27;5(7):e11803, Fig 1.

Immunocytochemistry/ Immunofluorescence - Anti-STAU2 antibody (ab60724)

ICC/IF image of ab60724 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab60724, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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) - STAU2 antibody (ab60724)

IHC image of ab60724 staining in human normal cervix 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 sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab60724, 5µg/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.