Anti-SHANK2 antibody [EPR24142-35]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR24142-35] (AB259966)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling SHANK2 with ab259966 at 1/5000 (0.0924 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Cytoplasmic stainig on mouse cerebrum (PMID : 29572432, 28179641). The section was incubated with ab259966 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SHANK2 antibody [EPR24142-35] (AB259966)

Immunofluorescence staining of SHANK2 using ab259966 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab259966 at 0.1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR24142-35] (AB259966)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling SHANK2 with ab259966 at 1/5000 (0.0924 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Positive staining on bile duct in rat liver (PMID : 14977424). The section was incubated with ab259966 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR24142-35] (AB259966)

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling SHANK2 with ab259966 at 1/5000 (0.0924 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Cytoplasmic stainig on mouse hippocampus (PMID : 27581745, 29572432, 28179641). The section was incubated with ab259966 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK2 antibody [EPR24142-35] (AB259966)

Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling SHANK2 with ab259966 at 1/5000 (0.0924 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Cytoplasmic stainig on rat hippocampus. The section was incubated with ab259966 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SHANK2 antibody [EPR24142-35] (AB259966)

Immunofluorescence staining of SHANK2 using ab259966 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab259966 at 1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• WB

Lab

Western blot - Anti-SHANK2 antibody [EPR24142-35] (AB259966)

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

Shank2 is expressed predominately in brain tissue what has described in the literature. (PMID : 9742101).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-SHANK2 antibody [EPR24142-35] (ab259966) at 1/1000 dilution

Lane 1:

Mouse hippocampus tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Lane 3:

Rat hippocampus tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 165 kDa

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