Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker

• WB

Supplier Data

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (AB109182)

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 (ab109182) 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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• WB

Unknown

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (AB109182)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (ab109182) at 1/5000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 105 kDa

Observed band size: 120 kDa

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• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (AB109182)

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.

• WB

Unknown

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (AB109182)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (ab109182) at 1/1000 dilution

All lanes:

Human cerebellum tissue lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 105 kDa

Observed band size: 120 kDa

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• WB

Unknown

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (AB109182)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (ab109182) at 1/5000 dilution

All lanes:

Human fetal brain tissue lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 105 kDa

Observed band size: 120 kDa

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• WB

Unknown

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (AB109182)

Secondary antibody - anti-rabbit HRP (ab6721)

All lanes:

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (ab109182) at 1/1000 dilution

All lanes:

Human fetal brain cell lysate at 10 µg

Predicted band size: 105 kDa

Observed band size: 120 kDa

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• WB

CiteAb

Western blot - Anti-NMDAR1 antibody [EPR2481(2)] - Neuronal Marker (AB109182)

NMDAR1 western blot using anti-NMDAR1 antibody [EPR2481(2)] ab109182. Publication image and figure legend from Li, J., Liu, Y., et al., 2019, Neural Plast, PubMed 30911292.

ab109182 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab109182 please see the product overview.

Hippocampal synaptic plasticity-associated proteins in diabetic rats and the effects of aerobic exercise. Levels of NMDAR1 (a), SYN (b), and Homer 1 (c) were significantly lower in diabetic rats. After aerobic exercise intervention, these protein levels were significantly increased. **p < 0.01, DM group vs. C group; ##p < 0.01, TDM group vs. DM group.

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