Anti-NMDAR1 antibody [N308/48] - Neuronal Marker (ab134308) at 1/1000 dilution + Rat brain membrane tissue lysate

Secondary
Goat anti-mouse IgG HRP

Predicted band size: 105 kDa

SK-N-BE cells labeling NMDAR1 using ab134308 at 1/100 dilution in ICC/IF. Cells were fixed using 4% formaldehyde for 15 minutes at room temperature. Incubated with primary antibody for 1 hour at room temperature. Secondary antibody was a goat anti-mouse ATTO 488 (green) at 1/100 dilution for 1 hour at room temperature. Counterstained with Phalloidin Texas Red F-actin stain. Nuclei were stained with DAPI (blue).

Overlay histogram showing SH-SY5Y cells stained with ab134308 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab134308, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.This antibody gave a positive result in 80% methanol (5 min) fixed SH-SY5Y cells used under the same conditions. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.