Anti-HCN4 antibody [EPR30362-531]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCN4 antibody [EPR30362-531] (AB327083)

Immunohistochemical analysis of paraffin-embedded Human thalamus tissue labeling HCN4 with ab327083 at 1/100 (5.04 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human thalamus. 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-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCN4 antibody [EPR30362-531] (AB327083)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human HCN4 expression vector containing a Myc-His tag cell pellet (B) HEK-293T transfected with a human HCN1 expression vector containing a Myc-His tag cell pellet (C) HEK-293T transfected with a human HCN2 expression vector containing a Myc-His tag cell pellet (D) HEK-293T transfected with a human HCN3 expression vector containing a His tag cell pellet (E) HEK-293T transfected with an empty vector containing a Myc-His tag cell pellet tissue labeling HCN4 with ab327083 at 1/5000 (0.101 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HEK-293T transfected with a human HCN4 expression vector containing a Myc-His tag cell pellet(image A), no staining on HEK-293T transfected with a human HCN1 expression vector containing a Myc-His tag cell pellet(image B), HEK-293T transfected with a human HCN2 expression vector containing a Myc-His tag cell pellet(image C), HEK-293T transfected with a human HCN3 expression vector containing a His tag cell pellet(image D) and HEK-293T transfected with an empty vector containing a Myc-His tag cell pellet(image E). 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-HCN4 antibody [EPR30362-531] (AB327083)

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

Negative control : liver, lung(PMID : 10228147)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 14657344

; PMID : 30877792).

The identity of the lower MW band at approximately 50 kDa is unknown.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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-HCN4 antibody [EPR30362-531] (ab327083) at 1/1000 dilution

Lane 1:

Mouse hypothalamus tissue lysate at 50 µg

Lane 2:

Mouse thalamus tissue lysate at 50 µg

Lane 3:

Mouse striatum tissue lysate at 50 µg

Lane 4:

Mouse lung tissue lysate at 50 µg

Lane 5:

Mouse liver tissue lysate at 50 µg

Lane 6:

Rat hypothalamus tissue lysate at 50 µg

Lane 7:

Rat striatum tissue lysate at 50 µg

Lane 8:

Rat lung tissue lysate at 50 µg

Lane 9:

Rat liver tissue lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 100-150 kDa,36 kDa

true

Exposure time: 81s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCN4 antibody [EPR30362-531] (AB327083)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HCN4 with ab327083 at 1/100 (5.04 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human liver. 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-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCN4 antibody [EPR30362-531] (AB327083)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling HCN4 with ab327083 at 1/100 (5.04 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human lung. 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