Anti-ASIC3 antibody [RM2049]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ASIC3 antibody [RM2049] (AB317460)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling ASIC3 with ab317460 at 1/4000 (0.124 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing positive staining in 293T cells (shown in green) transfected with a mouse ASIC3 expression vector containing a myc-His-tag® . The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ICC has not been tested with endogenous material.

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASIC3 antibody [RM2049] (AB317460)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a mouse ASIC3 expression vector containing a Myc-His tag. (B) HEK-293T transfected with empty vector containing a Myc-His tag. tissue labeling ASIC3 with ab317460 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HEK-293T transfected with a Myc-His-tagged mouse ASIC3 construct (image A). No staining on HEK-293T transfected with empty plasmid (image B). The section was incubated with ab317460 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

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Immunohistochemistry (Frozen sections) - Anti-ASIC3 antibody [RM2049] (AB317460)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat heart (fresh frozen) tissue labeling ASIC3 with ab317460 at 1/50 (9.92 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 heart (PMID : 9707631). The nuclear counterstain was DAPI (Blue). The section was incubated with ab317460 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-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ASIC3 antibody [RM2049] (AB317460)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse heart (fresh frozen) tissue labeling ASIC3 with ab317460 at 1/50 (9.92 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 heart. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317460 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-ASIC3 antibody [RM2049] (AB317460)

Immunohistochemical analysis of paraffin-embedded Mouse dorsal root ganglion tissue labeling ASIC3 with ab317460 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse dorsal root ganglion. The section was incubated with ab317460 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-ASIC3 antibody [RM2049] (AB317460)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse dorsal root ganglion (fresh frozen) tissue labeling ASIC3 with ab317460 at 1/50 (9.92 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-ASIC3 (ab317460, green) and anti-NeuN (ab190565, magenta) on mouse dorsal root ganglion.
Panel B : anti-ASIC3 stained on mouse dorsal root ganglion.
Panel C : anti-NeuN stained in neurons of mouse dorsal root ganglion.
The section was incubated in two rounds of staining : in the order of ab317460 and ab190565 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). 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-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ASIC3 antibody [RM2049] (AB317460)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat dorsal root ganglion (fresh frozen) tissue labeling ASIC3 with ab317460 at 1/50 (9.92 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-ASIC3 (ab317460, green) and anti-NeuN (ab190565, magenta) on rat dorsal root ganglion.
Panel B : anti-ASIC3 stained on rat dorsal root ganglion.
Panel C : anti-NeuN stained in neurons of rat dorsal root ganglion.
The section was incubated in two rounds of staining : in the order of ab317460 and ab190565 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). 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-ASIC3 antibody [RM2049] (AB317460)

Immunohistochemical analysis of paraffin-embedded Rat dorsal root ganglion tissue labeling ASIC3 with ab317460 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat dorsal root ganglion. The section was incubated with ab317460 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASIC3 antibody [RM2049] (AB317460)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling ASIC3 with ab317460 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse cardiac muscle. The section was incubated with ab317460 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASIC3 antibody [RM2049] (AB317460)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling ASIC3 with ab317460 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat cardiac muscle. The section was incubated with ab317460 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

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Western blot - Anti-ASIC3 antibody [RM2049] (AB317460)

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

Negative controls : hippocampus and testis.(PMID : 11872753)

The molecular weight is consistent with what has been described in the literature (PMID : 17012229)

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

All lanes:

Western blot - Anti-ASIC3 antibody [RM2049] (ab317460) at 1/1000 dilution

Lane 1:

Mouse dorsal ganglion tissue lysate at 40 µg

Lane 2:

Mouse hippocampus tissue lysate at 40 µg

Lane 3:

Mouse testis tissue lysate at 40 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: 55-60 kDa,124 kDa

false

Exposure time: 114s

• WB

Supplier Data

Western blot - Anti-ASIC3 antibody [RM2049] (AB317460)

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

Negative controls : hippocampus and testis.(PMID : 11872753)

The molecular weight is consistent with what has been described in the literature (PMID : 17012229)

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

All lanes:

Western blot - Anti-ASIC3 antibody [RM2049] (ab317460) at 1/1000 dilution

Lane 1:

Rat dorsal ganglion tissue lysate at 40 µg

Lane 2:

Rat hippocampus tissue lysate at 40 µg

Lane 3:

Rat testis tissue lysate at 40 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: 55-60 kDa,124 kDa

false

Exposure time: 70s