Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling HCN1 with ab252835 at 0.065, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on rat cerebrum tissue is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded rat retina tissue labeling HCN1 with ab252835 at 0.065µg/ml, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on rat retina tissue is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling HCN1 with ab252835 at 0.065µg/ml, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on mouse cerebrum tissue is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded mouse retina tissue labeling HCN1 with ab252835 at 0.065µg/ml, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on mouse retina tissue is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling HCN1 with ab252835 at 0.065µg/ml, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on human esophagus tissue is observed. The section was incubated with ab252835 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

All lanes : Anti-HCN1 antibody [7C3] (ab252835) at 0.326 µg/ml

Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 99 kDa


Exposure time: 3 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: 3 secs.

All lanes : Anti-HCN1 antibody [7C3] (ab252835) at 0.326 µg/ml

Lane 1 : Human brain tissue lysate
Lane 2 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 99 kDa


Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: 5 secs.

The negative control and the molecular weight of HCN1 observed is consistent with what has been described in the literature (PMID: 12786975).