ab9272 at a dilution of 1/1000, staining NCAM (Alexa 488 secondary at 1/2000) on rat brain tissue (30µm thick coronal sections) in free floating IHC (see protocol link for detailed description). Images showing hippocampal neuron body and processes: [A] 20x objective and [B] punctate cytoplasmic staining; 40x objective. No labeling observed following omission of primary antibody.

Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).

Immunofluorescence staining images of 9 day old zebrafish embryos.
ab9272 reacts with developing neuronal tissue (strong) and muscle. Frozen sample treated with Acetone:Methanol 1:1, antibody diluted 1/100 and incubated for 45 minutes at room temperature.

Immunofluorescence staining images of 9 day old zebrafish embryos.
ab9272 reacts with developing neuronal tissue (strong) and muscle. Frozen sample treated with Acetone:Methanol 1:1, antibody diluted 1/100 and incubated for 45 minutes at room temperature.

Immunofluorescence staining images of 9 day old zebrafish embryos.
ab9272 reacts with developing neuronal tissue (strong) and muscle. Frozen sample treated with Acetone:Methanol 1:1, antibody diluted 1/100 and incubated for 45 minutes at room temperature.

Immunostaining of the neural cell adhesion molecule (NCAM) in small cell human lung cancer cell line NCI-H82.

Overlay histogram showing SH-SY5Y cells stained with ab9272 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9272, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.