Anti-KCNQ2 antibody [EPR29531-83]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) and Differentiated SH-SY5Y cells labelling KCNQ2 with ab325971 at 1/50 (10.26 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in differentiated SH-SY5Y cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab78078 Anti-beta III Tubulin mouse monoclonal antibody - Neuronal Marker was used to counterstain tubulin at 1/1000 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh frozen) tissue labeling KCNQ2 with ab325971 at 1/50 (10.26 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green).

Confocal image showing positive staining on mouse hippocampus.
Panel A : merged staining of anti-KCNQ2 (ab325971, green), anti-NeuN (ab190565, magenta) on mouse hippocampus.
Panel B : anti-KCNQ2 stained on mouse hippocampus.
Panel C : anti-NeuN stained in neurons of mouse hippocampus.
The section was incubated in two rounds of staining : in the order of ab325971 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).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling KCNQ2 with ab325971 at 1/50 (10.26 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in mouse primary neural/glia cell (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325971 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab11267 at 1/500 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocampus (fresh frozen) tissue labeling KCNQ2 with ab325971 at 1/50 (10.26 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green).

Confocal image showing positive staining on rat hippocampus.
Panel A : merged staining of anti-KCNQ2 (ab325971, green), anti-NeuN (ab190565, magenta) on rat hippocampus.
Panel B : anti-KCNQ2 stained on rat hippocampus.
Panel C : anti-NeuN stained in neurons of rat hippocampus.
The section was incubated in two rounds of staining : in the order of ab325971 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).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse splenocytes cells labelling KCNQ2 with ab325971 at 1/50 (10.26 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Low expression : Confocal image showing no staining in mouse splenocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325971 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling KCNQ2 with ab325971 at 1/50 (10.26 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in rat primary neural/glia cell (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325971 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab11267 at 1/500 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunohistochemical analysis of paraffin-embedded Mouse brain tissue labeling KCNQ2 with ab325971 at 1/500 (1.026 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse brain.

The primary antibody was incubated 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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Rat splenocytes cells labelling KCNQ2 with ab325971 at 1/50 (10.26 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Low expression : Confocal image showing no staining in rat splenocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325971 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunohistochemical analysis of paraffin-embedded Rat brain tissue labeling KCNQ2 with ab325971 at 1/500 (1.026 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in rat brain.

The primary antibody was incubated 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

Lab

Western blot - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 14762142).

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

The identity of the bands between 25 kDa and 37 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-KCNQ2 antibody [EPR29531-83] (ab325971) at 1/1000 dilution

Lane 1:

Untreated SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

SH-SY5Y treated with 10 uM retinoic acid for 7 days, 50ng/ml BDNF was then added for additional 7 days whole cell lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 100 kDa,75 kDa,36 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

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

Negative control : heart, spleen (PMID : 9872318).

Low expression : spinal cord (PMID : 14762142, 9872318).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 14762142).

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-KCNQ2 antibody [EPR29531-83] (ab325971)

Lane 1:

Mouse hippocampus tissue lysate at 20 µg

Lane 2:

Mouse cerebral cortex tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse heart tissue lysate at 20 µg

Lane 5:

Mouse spleen tissue lysate at 20 µg

Lane 6:

Mouse spinal cord tissue lysate at 20 µg

Lane 7:

Rat hippocampus tissue lysate at 20 µg

Lane 8:

Rat heart tissue lysate at 20 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: 100 kDa,75 kDa,36 kDa

false

Exposure time: 48s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling KCNQ2 with ab325971 at 1/500 (1.026 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in mouse cardiac muscle.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling KCNQ2 with ab325971 at 1/500 (1.026 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in rat cardiac muscle.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunohistochemical analysis of paraffin-embedded
(A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a mouse KCNQ2 expression vector containing a Myc-His tag.
(B) HEK-293T transfected with a mouse KCNQ1 expression vector containing a Myc-His tag.
(C) HEK-293T transfected with a mouse KCNQ3 expression vector containing a Myc-His tag.
(D) HEK-293T transfected with a mouse KCNQ4 expression vector containing a Myc-His tag.
(E) HEK-293T transfected with a mouse KCNQ5 expression vector containing a Myc-His tag.
(F) HEK-293T transfected with empty vector containing a Myc-His tag.
labeling KCNQ2 with ab325971 at 1/20000 (0.026 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) HEK-293T transfected with a mouse KCNQ2 expression vector containing a Myc-His tag, and no signal in (B) HEK-293T transfected with a mouse KCNQ1 expression vector containing a Myc-His tag, (C) HEK-293T transfected with a mouse KCNQ3 expression vector containing a Myc-His tag, (D) HEK-293T transfected with a mouse KCNQ4 expression vector containing a Myc-His tag, (E) HEK-293T transfected with a mouse KCNQ5 expression vector containing a Myc-His tag, (F) HEK-293T transfected with empty vector containing a Myc-His tag.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Frozen sections) - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse heart (fresh frozen) tissue labeling KCNQ2 with ab325971 at 1/50 (10.26 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green).

Negative control : confocal image showing no staining on mouse heart. The nuclear counterstain was DAPI (Blue). The section was incubated with ab325971 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) preadsorbed at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-KCNQ2 antibody [EPR29531-83] (AB325971)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat heart (fresh frozen) tissue labeling KCNQ2 with ab325971 at 1/50 (10.26 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green).

Negative control : confocal image showing no staining on rat heart. The nuclear counterstain was DAPI (Blue). The section was incubated with ab325971 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) preadsorbed at 1/1000 dilution.