Recombinant Anti-Kv4.2/KCND2 antibody [EPR26384-89] (ab307710)

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

This antibody does not cross-react with mouse KCND1 and KCND3.

In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution.

Exposure time: 10 seconds

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

Negative control:  liver(Human Protein Atlas Database), bladder (PMID:12575952).

The molecular weight observed is consistent with what has been described in the literature (PMID:15454437).

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

Exposure time: 26 seconds

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling Kv4.2/KCND2 with ab307710 at 1/2000 (0.266 μg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

Cytoplasmic staining on rat cerebellum (PMID: 32899153).

The section was incubated with ab307710 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 LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh) tissue labeling Kv4.2/KCND2 with ab307710 at 1/100 (5.32 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green).

Negative control: confocal image showing no staining on rat liver (PMID: 10729221).

The nuclear counterstain was DAPI (Blue). The section was incubated with ab307710 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). The nuclear counterstain was DAPI (Blue).

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

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat dorsal root ganglion (fresh) tissue labeling Kv4.2/KCND2 with ab307710 at 1/100 (5.32 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green).

Panel A: merged staining of anti-Kv4.2 (ab307710, green) and anti-NeuN (ab190565, red) on rat dorsal root ganglion.

Panel B: anti-Kv4.2 stained on the rat dorsal root ganglion.

Panel C: anti-NeuN stained in neurons of rat dorsal root ganglion.

Negative control: dorsal root ganglion (PMID: 19668710).

The nuclear counterstain was DAPI (Blue). The section was incubated in two rounds of staining: in the order of ab307710 and ab190565 for 1 hr 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). The nuclear counterstain was DAPI (Blue).

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

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebellum (fresh) tissue labeling Kv4.2/KCND2 with ab307710 at 1/100 (5.32 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green).

Panel A: merged staining of anti-Kv4.2 (ab307710, green) and anti-NeuN (ab190565, red) on rat cerebellum.

Panel B: anti-Kv4.2 stained on the rat cerebellum.

Panel C: anti-NeuN stained in neurons of rat cerebellum. The nuclear counterstain was DAPI (Blue). The section was incubated in two rounds of staining: in the order of ab307710 and ab190565 for 1 hr 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). The nuclear counterstain was DAPI (Blue).

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

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh) tissue labeling Kv4.2/KCND2 with ab307710 at 1/100 (5.32 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green).

Negative control: confocal image showing no staining on mouse liver (PMID: 10729221).

The nuclear counterstain was DAPI (Blue). The section was incubated with ab307710 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). The nuclear counterstain was DAPI (Blue).

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

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse dorsal root ganglion (fresh) tissue labeling Kv4.2/KCND2 with ab307710 at 1/100 (5.32 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green).

Panel A: merged staining of anti-Kv4.2 (ab307710, green) and anti-NeuN (ab190565, red) on mouse dorsal root ganglion.

Panel B: anti-Kv4.2 stained on the mouse dorsal root ganglion.

Panel C: anti-NeuN stained in neurons of mouse dorsal root ganglion.

Negative control: dorsal root ganglion (PMID: 19668710).The nuclear counterstain was DAPI (Blue).

The section was incubated in two rounds of staining: in the order of ab307710 and ab190565 for 1 hr 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). The nuclear counterstain was DAPI (Blue).

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

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum (fresh) tissue labeling Kv4.2/KCND2 with ab307710 at 1/100 (5.32 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green).

Panel A: merged staining of anti-Kv4.2 (ab307710, green) and anti-NeuN (ab190565, red) on mouse cerebellum.

Panel B: anti-Kv4.2 stained on the mouse cerebellum.

Panel C: anti-NeuN stained in neurons of mouse cerebellum. The nuclear counterstain was DAPI (Blue). The section was incubated in two rounds of staining: in the order of ab307710 and ab190565 for 1 hr 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). The nuclear counterstain was DAPI (Blue).

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

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat primary neuron cells cells labelling Kv4.2/KCND2 with ab307710 at 1/50 dilution (1μg)/ 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). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cells cells labelling Kv4.2/KCND2 with ab307710 at 1/50 dilution (1μg)/ 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). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

Kv4.2/KCND2 was immunoprecipitated from 0.35 mg Rat cerebellum tissue lysate 10 μg with ab307710 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307710 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Rat cerebellum tissue lysate 10 μg

Lane 2: ab307710 IP in Rat cerebellum tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab307710 in rat cerebellum tissue lysate

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

Exposure time: 8 seconds

Kv4.2/KCND2 was immunoprecipitated from 0.35 mg Mouse cerebellum tissue lysate 10 μg with ab307710 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307710 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse cerebellum tissue lysate 10 μg

Lane 2: ab307710 IP in Mouse cerebellum tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab307710 in mouse cerebellum tissue lysate

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

Exposure time: 8 seconds

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Kv4.2/KCND2 with ab307710 at 1/2000 (0.266 μg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

Negative control: no staining on rat liver.

The section was incubated with ab307710 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 LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Kv4.2/KCND2 with ab307710 at 1/2000 (0.266 μg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

Negative control: no staining on mouse liver (PMID: 16293790).

The section was incubated with ab307710 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 LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling Kv4.2/KCND2 with ab307710 at 1/2000 (0.266 μg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

Cytoplasmic staining on mouse cerebellum (PMID: 17122039).

The section was incubated with ab307710 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 LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins