Immunocytochemistry/Immunofluorescence analysis of human neuroblastoma cells (SH-SY5Y), fixed in 4% PFA for 15 min, using Anti-HCN1 Antibody [S70] (A304768), at 1:100 for overnight at 4°C with slow rocking. The secondary antibody used was AlexaFluor 488 at 1:1,000 for 1 hour at room temperature. Counterstain: Phalloidin-iFluor 647 (red) F-Actin stain; Hoechst (blue) nuclear stain at 1:800, 1.6mM for 20 minutes at room temperature.(A) Hoechst (blue) nuclear stain. (B) Phalloidin-iFluor 647 (red) F-Actin stain. (C) HCN1 Antibody (D) Composite.
Immunohistochemistry analysis of mouse frozen brain section, fixed in 10% formalin solution for 12-24 hours at room temperature. The Primary Antibody used was Anti-HCN1 Antibody [S70] (A304768) at 1:1,000 for 1 hour at room temperature. The secondary antibody used was HRP/DAB Detection System: Biotinylated Goat Anti-Mouse, Streptavidin Peroxidase, DAB Chromogen (brown) for 30 minutes at room temperature. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 250-500 µl for 5 minutes at room temperature.
Immunohistochemistry analysis of mouse frozen brain section, fixed in 10% formalin solution for 12-24 hours at room temperature. The Primary Antibody used was Anti-HCN1 Antibody [S70] (A304768) at 1:1,000 for 1 hour at room temperature. The secondary antibody used was HRP/DAB Detection System: Biotinylated Goat Anti-Mouse, Streptavidin Peroxidase, DAB Chromogen (brown) for 30 minutes at room temperature. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 250-500 µl for 5 minutes at room temperature. Localization: Neurons.
Immunohistochemistry analysis of human hippocampus, fixed in Bouin's fixative solution and paraffin-embedded. The Primary Antibody used was Anti-HCN1 Antibody [S70] (A304768) at 1:1,000 for 1 hour at room temperature. The secondary antibody used was FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at room temperature.
Western blot analysis of rat brain membrane lysate showing detection of HCN1 protein using Anti-HCN1 Antibody [S70] (A304768) at 1:1,000 for 2 hours at room temperature. Load: 15 µg. Block: 1.5% BSA for 30 minutes at room temperature. The secondary antibody used was Sheep Anti-Mouse IgG: HRP for 1 hour at room temperature.
Immunohistochemistry analysis of mouse cerebellum, fixed in 10% formalin solution for 12-24 hours at room temperature. The Primary Antibody used was Anti-HCN1 Antibody [S70] (A304768) at 1:1,000 for 1 hour at room temperature. The secondary antibody used was HRP/DAB Detection System: Biotinylated Goat Anti-Mouse, Streptavidin Peroxidase, DAB Chromogen (brown) for 30 minutes at room temperature. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 250-500 µl for 5 minutes at room temperature. Localization: Cytoplasmic staining of Purkinje cells.
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