Anti-HCN2 antibody [EPR26389-129]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling HCN2 with ab313873 at 1/100 (5.05 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human skeletal muscle. The section was incubated with ab313873 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-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling HCN2 with ab313873 at 1/100 (5.05 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab313873 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-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling HCN2 with ab313873 at 1/100 (5.05 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on rat skeletal muscle. The section was incubated with ab313873 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-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling HCN2 with ab313873 at 1/100 (5.05 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse skeletal muscle. The section was incubated with ab313873 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-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling HCN2 with ab313873 at 1/100 (5.05 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum (PMID : 14991560). The section was incubated with ab313873 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-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling HCN2 with ab313873 at 1/100 (5.05 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cerebrum (PMID : 14991560). The section was incubated with ab313873 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-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum (fresh) tissue labeling HCN2 with ab313873 at 1/50 (10.1 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/ml dilution (Green). Confocal image showing positive staining on rat cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab313873 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). is observed. 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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat skeletal muscle (fresh) tissue labeling HCN2 with ab313873 at 1/50 (10.1 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 skeletal muscle (PMID : 10228147). The nuclear counterstain was DAPI (Blue). The section was incubated with ab313873 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). is observed. 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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skeletal muscle (fresh) tissue labeling HCN2 with ab313873 at 1/50 (10.1 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 skeletal muscle (PMID : 10228147). The nuclear counterstain was DAPI (Blue). The section was incubated with ab313873 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). is observed. 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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-HCN2 antibody [EPR26389-129] (AB313873)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh) tissue labeling HCN2 with ab313873 at 1/50 (10.1 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/ml dilution (Green). Confocal image showing positive staining on mouse cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab313873 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). is observed. 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.

• IP

Supplier Data

Immunoprecipitation - Anti-HCN2 antibody [EPR26389-129] (AB313873)

HCN2 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab313873 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313873 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse brain tissue lysate Lane 2 : ab313873 IP in Mouse brain tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313873 in mouse brain tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 93 seconds

All lanes:

Immunoprecipitation - Anti-HCN2 antibody [EPR26389-129] (ab313873) at 1/30 dilution

All lanes:

Mouse brain tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 93s

• WB

Lab

Western blot - Anti-HCN2 antibody [EPR26389-129] (AB313873)

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

Negative control : lung (PMID : 10228147).

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

All lanes:

Western blot - Anti-HCN2 antibody [EPR26389-129] (ab313873) at 1/1000 dilution

Lanes 1 and 5:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lanes 3 and 6:

Rat brain tissue lysate at 20 µg

Lane 4:

Rat lung 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,110 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-HCN2 antibody [EPR26389-129] (AB313873)

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

Negative control : liver (PMID : 10228147, PMID : 9630217); spleen, testis (PMID : 9630217).

This antibody detects bands at ~100 kDa and ~110 kDa representing the non-glycosylated and glycosylated form of HCN2(PMID : 19236845)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HCN2 antibody [EPR26389-129] (ab313873) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Lane 4:

Rat spleen 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: 97 kDa

true

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-HCN2 antibody [EPR26389-129] (AB313873)

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

Negative control : lung, skeletal muscle (PMID : 10228147, PMID : 9630217); testis (PMID : 9630217).

Bands above 250 kDa are aggregates of HCN2.

This antibody detects bands at ~100 kDa and ~110 kDa representing the non-glycosylated and glycosylated form of HCN2(PMID : 19236845)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HCN2 antibody [EPR26389-129] (ab313873) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse testis 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: 110 kDa,100 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-HCN2 antibody [EPR26389-129] (AB313873)

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

Negative control : skeletal muscle (PMID : 10228147, PMID : 9630217)

Bands above 250 kDa are aggregates of HCN2.

This antibody detects bands at ~100 kDa and ~110 kDa representing the non-glycosylated and glycosylated form of HCN2(PMID : 19236845)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HCN2 antibody [EPR26389-129] (ab313873) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat skeletal muscle 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: 110 kDa,100 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-HCN2 antibody [EPR26389-129] (AB313873)

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

Bands above 250 kDa are aggregates of HCN2.

This antibody detects bands at ~100 kDa and ~110 kDa representing the non-glycosylated and glycosylated form of HCN2(PMID : 19236845).

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

All lanes:

Western blot - Anti-HCN2 antibody [EPR26389-129] (ab313873) at 1/1000 dilution

Lane 1:

Mouse hippocampus tissue lysate at 20 µg

Lane 2:

Untreated mouse brain tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate treated with Protein treated with Peptide:N-glycosidase F (PNGase F) 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,110 kDa

true

Exposure time: 3min