Anti-MAP2 antibody - Neuronal Marker (ab32454)

  Immunocytochemistry/ Immunofluorescence - MAP2 antibody (ab32454)

ICC/IF image of ab32454 stained rat PC12 cells. The cells were PFA fixed (10 min), permabilised in PBS-T (20 min) and incubated with the antibody (ab32454, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - MAP2 antibody - Neuronal Marker (ab32454)

IHC image of MAP2 staining in human cerebral cortex FFPE 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 ab32454, 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.

  Western blot - MAP2 antibody - Neuronal Marker (ab32454)

All lanes : Anti-MAP2 antibody - Neuronal Marker (ab32454) at 1 µg/ml

Lane 1 : Brain (Mouse) Tissue Lysate - normal tissue, 0 days old (ab7188)
Lane 2 : Brain (Rat) Tissue Lysate

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 70, 199, 280 kDa
Observed band size : 260,280 kDa (why is the actual band size different from the predicted?)
Additional bands at : 110 kDa,199 kDa,65 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 3 minutes