Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized T-47D (human ductal breast epithelial tumor epithelial cell) (Right) / 293T (human embryonic kidney epithelial cell) (Left) cells labelling SHANK2 with ab317606 at 1/500 dilution (0.1ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : 293T(PMID : 32661924)

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh frozen) tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining on mouse hippocampus. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317606 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse hippocampus. The section was incubated with ab317606 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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat primary neuron cells labelling SHANK2 with ab317606 at 1/500 dilution (0.1ug)/right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on cholangiocytes of mouse liver. The section was incubated with ab317606 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-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocampus (fresh frozen) tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining on rat hippocampus. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317606 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum. The section was incubated with ab317606 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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cells labelling SHANK2 with ab317606 at 1/500 dilution (0.1ug)/right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on cholangiocytes of rat liver. The section was incubated with ab317606 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

Supplier Data

Western blot - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Negative control : skeletal muscle (PMID : 10506216; PMID : 22346768).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29572432). The bands beneath the target band (180-200 kDa) are likely to be degraded target fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SHANK2 antibody [EPR26549-331] (<a href='/en-us/products/primary-antibodies/shank2-antibody-epr26549-331-ab317606'>ab317606</a>) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg with NFDM/TBST

Lane 2:

Human skeletal muscle tissue lysate at 20 µg with NFDM/TBST

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 1/200000 dilution

Observed band size: 200 kDa,180 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Low expression : 293T (PMID : 32661924).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29572432). The bands beneath the target band (180-270 kDa) are likely to be degraded target fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SHANK2 antibody [EPR26549-331] (<a href='/en-us/products/primary-antibodies/shank2-antibody-epr26549-331-ab317606'>ab317606</a>) at 1/1000 dilution

Lane 1:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

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: 180-270 kDa,124 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Negative control : skeletal muscle (PMID : 10506216; PMID : 22346768).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29572432). The bands beneath the target band (180-270 kDa) are likely to be degraded target fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SHANK2 antibody [EPR26549-331] (<a href='/en-us/products/primary-antibodies/shank2-antibody-epr26549-331-ab317606'>ab317606</a>) at 1/1000 dilution

Lane 1:

Mouse hippocampus tissue lysate at 20 µg with NFDM/TBST

Lane 2:

Mouse cerebellum tissue lysate at 20 µg with NFDM/TBST

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg with NFDM/TBST

Lane 4:

Rat hippocampus tissue lysate at 20 µg with NFDM/TBST

Lane 5:

Rat cerebellum tissue lysate at 20 µg with NFDM/TBST

Lane 6:

Rat skeletal muscle tissue lysate at 20 µg with NFDM/TBST

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: 180-270 kDa,180 kDa,124 kDa

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat skeletal muscle. The section was incubated with ab317606 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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse spleen cell cells labelling SHANK2 with ab317606 at 1/500 dilution (0.1ug)/right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : mouse spleen cell.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skeletal muscle (fresh frozen) tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Negative control : confocal image showing no staining on mouse skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317606 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse skeletal muscle. The section was incubated with ab317606 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-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat skeletal muscle (fresh frozen) tissue labeling SHANK2 with ab317606 at 1/100 (5.21 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Negative control : confocal image showing no staining on rat skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317606 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat spleen cell cells labelling SHANK2 with ab317606 at 1/500 dilution (0.1ug)/right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : rat spleen cell.

• WB

Supplier Data

Western blot - Anti-SHANK2 antibody [EPR26549-331] - BSA and Azide free (AB317607)

This data was developed using ab317606, the same antibody clone in a different buffer formulation.

This antibody does not cross-react with human SHANK1 and SHANK3.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-SHANK2 antibody [EPR26549-331] (<a href='/en-us/products/primary-antibodies/shank2-antibody-epr26549-331-ab317606'>ab317606</a>) at 1/1000 dilution

Lane 1:

His-tagged human SHANK2 recombinant fragment at 10 ng with NFDM/TBST

Lane 2:

His-tagged human SHANK1 recombinant fragment at 10 ng with NFDM/TBST

Lane 3:

His-tagged human SHANK3 recombinant fragment at 10 ng with NFDM/TBST

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 1/100000 dilution

Observed band size: 26 kDa

false

Exposure time: 70s