Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] - BSA and Azide free (AB240061)

Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Ionotropic Glutamate receptor 2 (red) with ab133477 at a 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133477).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] - BSA and Azide free (AB240061)

This data was developed using ab133477, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of Neuro-2a (Mouse neuroblastoma neuroblast) cells labeling Ionotropic Glutamate receptor 2 with purified ab133477 at 1/70 dilution (10 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-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.

• WB

Lab

Western blot - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] - BSA and Azide free (AB240061)

This data was developed using ab133477, the same antibody clone in a different buffer formulation.

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

Exposure time : ab133477 : 20s, ab181602 : 60s.

All lanes:

Western blot - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] (<a href='/ko/products/primary-antibodies/ionotropic-glutamate-receptor-2-antibody-epr5032-ab133477'>ab133477</a>) at 1/5000 dilution

Lane 1:

Mouse brain tissue whole cell lysate at 20 µg

Lane 2:

Rat brain tissue whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] - BSA and Azide free (AB240061)

This data was developed using ab133477, the same antibody clone in a different buffer formulation.

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

In Western blot, Anti-Myc tag antibody [9E10] - Loading Control (ab32) staining at 1/10000 dilution.

This antibody does not cross-react with Human GRIA1, GRIA3 and GRIA4.

Exposure time : ab133477 : 20s, ab32 : 80s.

All lanes:

Western blot - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] (<a href='/ko/products/primary-antibodies/ionotropic-glutamate-receptor-2-antibody-epr5032-ab133477'>ab133477</a>) at 1/5000 dilution

Lane 1:

293T transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T transfected with Human fragment GRIA2 expression vector containing a c-term myc-His-tag®, whole cell lysate at 20 µg

Lane 3:

293T transfected with Human full length GRIA3 expression vector containing a n-term myc-His-tag®, whole cell lysate at 20 µg

Lane 4:

293T transfected with human full length GRIA4 expression vector containing a c-term myc-His-tag®, whole cell lysate at 20 µg

Lane 5:

Recombinant Human full length GRIA1 Protein containing a C-Myc/DDK tag at 100 ng

Secondary

Lanes 1 - 5:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lanes 1 - 5:

Goat anti-mouse IgG (H+L), Peroxidase conjugated at 1/10000 dilution

Observed band size: 100 kDa

false

• WB

Unknown

Western blot - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] - BSA and Azide free (AB240061)

All lanes:

Western blot - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] (<a href='/ko/products/primary-antibodies/ionotropic-glutamate-receptor-2-antibody-epr5032-ab133477'>ab133477</a>) at 1/5000 dilution

Lane 1:

Human hypothalamus lysate at 15 µg

Lane 2:

Human midbrain lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 98 kDa

false

• WB

Unknown

Western blot - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] - BSA and Azide free (AB240061)

All lanes:

Western blot - Anti-Ionotropic Glutamate receptor 2 antibody [EPR5032] - BSA and Azide free (ab240061) at 1/1000 dilution

Lane 1:

SK-N-BE (2) (Human neuroblastoma neuroblast) whole cell lysate

Lane 2:

Mouse brain lysate boiled

Lane 3:

Mouse brain lysate unboiled

Lane 4:

Rat brain lysate boiled

Lane 5:

Rat brain lysate unboiled

Secondary

All lanes:

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

Predicted band size: 98 kDa

false