Anti-HCN1 antibody [S70]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCN1 antibody [S70] (AB84816)

Immunohistochemical analysis of paraffin-embedded (Bouin's Fixative - picric acid, acetic acid and formaldehyde) human hippocampus tissue labelling HCN1 with ab84816 at 1/1000 dilution, for 1 hour at RT followed by secondary antibody Goat Anti-Mouse (FITC) at 1/50 dilution, for 1 hour at RT. Membrane and cytosolic staining on human hippocampus tissue is observed.

• Flow Cyt

Unknown

Flow Cytometry - Anti-HCN1 antibody [S70] (AB84816)

Overlay histogram showing SH-SY5Y cells stained with ab84816 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab84816, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween for 20 min used under the same conditions.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-HCN1 antibody [S70] (AB84816)

Immunohistochemistry analysis of 10% formalin-fixed mouse frozen brain section labelling HCN1 with ab84816 at 1/1000 dilution for 1 hour at room temperature, followed by biotinylated Goat Anti-Mouse antibody (brown) for 30 minutes at room temperature. Counterstain : Hematoxylin (purple/blue) nuclear stain at 250-500 μl for 5 minutes at room temperature. HRP/DAB detection.

• WB

Unknown

Western blot - Anti-HCN1 antibody [S70] (AB84816)

All lanes:

Western blot - Anti-HCN1 antibody [S70] (ab84816) at 1/1000 dilution

All lanes:

Rat brain membrane lysates at 15 µg

Secondary

All lanes:

HRP-Sheep Anti-Mouse IgG antibody

Predicted band size: 99 kDa

Observed band size: 99 kDa

false

• IHC

CiteAb

Immunohistochemistry - Anti-HCN1 antibody [S70] (AB84816)

Immunohistochemistry-immunofluorescence using Anti-HCN1 antibody [S70], ab84816. Publication image from Kaestner, K. H. et al., 2016, Nat Commun, 27399229. Legend direct from paper.

β cell heterogeneity in normal pancreatic tissue sections.(a) Human pancreas co-labelled with antibodies recognizing CD9 and proinsulin. Most proinsulin+ β cells were CD9− (hollow arrows) but a subset was CD9+ (solid arrows). (b) Tissue co-labelled with antibodies recognizing ST8SIA1 and proinsulin. Most proinsulin+ β cells were ST8SIA1− (hollow arrows) but a subset was ST8SIA1+ (solid arrows). (c) Compound labelling of proinsulin, CD9 and ST8SIA1 reveals proinsulin+ cells expressing every combination of the other markers; examples of β1 (CD9−ST8SIA1−), β2 (CD9+ST8SIA1−), β3 (CD9−ST8SIA1+) and β4 (CD9+ST8SIA1+) cells are indicated. Note that both CD9 and ST8SIA1 are found on proinsulin− (non-β) cell types as well, as shown in Supplementary Fig. 1. (d) HCN1 was detected in a subset of proinsulin+ β cells; examples of high/positive and low/negative cells are indicated with solid and hollow arrows, respectively. Tissues illustrated are 5 µm cryosections of normal human pancreas. Scale bar, 25 µm.

• IHC

CiteAb

Immunohistochemistry - Anti-HCN1 antibody [S70] (AB84816)

Immunohistochemistry-immunofluorescence using Anti-HCN1 antibody [S70], ab84816. Publication image from Kaestner, K. H. et al., 2016, Nat Commun, 27399229. Legend direct from paper.

β cell heterogeneity in normal pancreatic tissue sections.(a) Human pancreas co-labelled with antibodies recognizing CD9 and proinsulin. Most proinsulin+ β cells were CD9− (hollow arrows) but a subset was CD9+ (solid arrows). (b) Tissue co-labelled with antibodies recognizing ST8SIA1 and proinsulin. Most proinsulin+ β cells were ST8SIA1− (hollow arrows) but a subset was ST8SIA1+ (solid arrows). (c) Compound labelling of proinsulin, CD9 and ST8SIA1 reveals proinsulin+ cells expressing every combination of the other markers; examples of β1 (CD9−ST8SIA1−), β2 (CD9+ST8SIA1−), β3 (CD9−ST8SIA1+) and β4 (CD9+ST8SIA1+) cells are indicated. Note that both CD9 and ST8SIA1 are found on proinsulin− (non-β) cell types as well, as shown in Supplementary Fig. 1. (d) HCN1 was detected in a subset of proinsulin+ β cells; examples of high/positive and low/negative cells are indicated with solid and hollow arrows, respectively. Tissues illustrated are 5 µm cryosections of normal human pancreas. Scale bar, 25 µm.