Anti-GluN2D antibody [EPR30355-38]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GluN2D antibody [EPR30355-38] (AB326329)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling GluN2D with ab326329 at 1/500 (1.012 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum (PMID : 25164652; PMID : 10216082). The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GluN2D antibody [EPR30355-38] (AB326329)

Immunohistochemical analysis of paraffin-embedded Mouse spinal cord tissue labeling GluN2D with ab326329 at 1/500 (1.012 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spinal cord. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Lab

Western blot - Anti-GluN2D antibody [EPR30355-38] (AB326329)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : colon, skeletal muscle, liver, spleen.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-GluN2D antibody [EPR30355-38] (ab326329) at 1/1000 dilution

Lane 1:

Mouse P5 brain tissue lysate at 20 µg

Lane 2:

Mouse colon tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Lane 5:

Mouse spleen tissue lysate at 20 µg

Lane 6:

Mouse cerebellum tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 165 kDa,36 kDa

false

Exposure time: 125s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GluN2D antibody [EPR30355-38] (AB326329)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling GluN2D with ab326329 at 1/500 (1.012 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse skeletal muscle. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Lab

Western blot - Anti-GluN2D antibody [EPR30355-38] (AB326329)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with overexpressed human GluN2A, GluN2B or GluN2C by western blot.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (165KDa).

All lanes:

Western blot - Anti-GluN2D antibody [EPR30355-38] (ab326329) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a human GluN2A expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 3:

293T cells transfected with a human GluN2B expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 4:

293T cells transfected with a human GluN2C expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 5:

293T cells transfected with a human GluN2D expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 165 kDa,36 kDa

false

Exposure time: 125s