Anti-NMDAR1 antibody [EPR2481(2)] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - BSA and Azide free (AB239949)

Western blot : Anti-GRIN1 antibody [EPR2481(2)] (ab109182) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab109182 was shown to bind specifically to GRIN1. A band was observed at 120 kDa in wild-type Neuro-2a cell lysates with no signal observed at this size in GRIN1 knockout cell line ab281960 (knockout cell lysate ab282987). To generate this image, wild-type and GRIN1 knockout Neuro-2a cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/nmdar1-antibody-epr24812-neuronal-marker-ab109182'>ab109182</a>) at 1/1000 dilution

Lane 1:

Wild-type Neuro-2a cell lysate at 20 µg

Lane 2:

GRIN1 knockout Neuro-2a cell lysate at 20 µg

Lane 3:

SH-SY5Y UNBOILED cell lysate at 20 µg

Lane 4:

THP-1 UNBOILED cell lysate at 20 µg

Lane 5:

Human Brain UNBOILED cell lysate at 20 µg

Lane 6:

Human Liver UNBOILED cell lysate at 20 µg

Lane 7:

Mouse Brain UNBOILED cell lysate at 20 µg

Lane 8:

Mouse Liver UNBOILED cell lysate at 20 µg

Observed band size: 120 kDa

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• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NMDAR1 antibody [EPR2481(2)] - BSA and Azide free (AB239949)

Immunocytochemistry/ Immunofluorescence analysis of mouse primary neuron cells labeling NMDAR1 with purified ab109182 at 1/50 (9.5μg/mL). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109182).