Recombinant Anti-ACTH antibody [57] - N-terminal (ab233954)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human pituitary gland labelling ACTH with ab233954 at 1/5000 dilution (0.214 μg/ml) (B), Luteinizing Hormone beta with ab221494 at 1/4000 dilution (0.258 μg/ml) (C) and Growth Hormone with ab155276 at 1/5000 dilution (0.044 μg/ml) (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A: merged staining of anti-Growth Hormone (gray; Opal™690), anti-ACTH (green; Opal™520) and anti-Luteinizing Hormone beta (red; Opal™570) on human pituitary gland.
Panel B: anti-ACTH stained on corticotrophs.
Panel C: anti-Luteinizing Hormone beta stained on gonadotrophs.
Panel D: anti-Growth Hormone stained on somatotrophs.

The section was incubated in three rounds of staining: in the order of ab155276, ab233954, and ab221494 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Fluorescence multiplex immunohistochemical analysis of human pituitary gland tissue (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-ACTH stained on corticotrophs (ab233954; green; Opal™520) at 1:5000 (0.214 μg/ml) [Panel B], anti-TSH beta stained on thyrotrophs (ab155958; red; Opal™570) at 1:500 (1.728 μg/ml) [Panel C], and anti-Growth Hormone stained on somatotrophs (ab155276; gray; Opal™690) at 1:5000 (0.044 μg/ml) [Panel D] on human pituitary gland. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of ab155276, ab233954, and ab155958 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used. 

Immunohistochemical analysis of Human pituatary gland tissue stained for ACTH using ab233954 at 1/5000 dilution. Positive staining on human pituitary gland. The section was incubated with ab233954 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Immunohistochemical analysis of Mouse pituatary gland tissue stained for ACTH using ab233954 at 1/20000 dilution. Positive staining on mouse pituitary gland. The section was incubated with ab233954 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Immunohistochemical analysis of Rat pituatary gland tissue stained for ACTH using ab233954 at 1/20000 dilution. Positive staining on rat pituitary gland. The section was incubated with ab233954 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.