Journal: Journal of Histochemistry and Cytochemistry

Article Title: Immunocytochemical Profiling of Cultured Mouse Primary Retinal Cells

doi: 10.1369/0022155416689675

Figure Lengend Snippet: Secondary Antibody List.

Article Snippet: Donkey , Anti-sheep , FITC , 1:200 , Jackson ImmunoResearch Laboratories, Inc., West Grove, PA , 713-095-147.

Techniques:

Journal: Frontiers in Neuroanatomy

Article Title: Localization of Vasoactive Intestinal Polypeptide Receptor 1 (VPAC1) in Hypothalamic Neuroendocrine Oxytocin Neurons; A Potential Role in Circadian Prolactin Secretion

doi: 10.3389/fnana.2020.579466

Figure Lengend Snippet: Secondary antibodies.

Article Snippet: After 15-min fixation in 4% PFA, hybridization overnight, and washing, slides were incubated with a sheep anti-digoxigenin antibody (Roche, code: 11 214 667 001; diluted 1:2,000) followed by washing and incubation with a biotinylated donkey anti–sheep antibody (Jackson ImmunoResearch code; 713-066-147, diluted 1:800), washed followed by incubation in ABC reagent (Vectastain PK6100) biotinylated tyramide (Perkin Elmer SAT 700, diluted 1:50) and finally in streptavidin conjugated Alexa Fluor 488 (Jackson ImmunoResearch; code 016-540-084, diluted 1:500).

Techniques:

Journal: Frontiers in Neuroanatomy

Article Title: Localization of Vasoactive Intestinal Polypeptide Receptor 1 (VPAC1) in Hypothalamic Neuroendocrine Oxytocin Neurons; A Potential Role in Circadian Prolactin Secretion

doi: 10.3389/fnana.2020.579466

Figure Lengend Snippet: Distribution of VPAC2 (green) and vasoactive intestinal polypeptide (VIP; red) immunoreactivity (A–C) and distribution of VPAC1 (green) and VIP (red) immunoreactivity (D–F) in the mid and rostral part of the suprachiasmatic nucleus (SCN) and paraventricular nucleus (PVN) in a wild-type mouse. Images (A–E,G,H) represent maximal projections of 14 digital sections in a Z-stack ( Z = 2 μm). Image I represents a 3D reconstruction of a Z-stack ( Z = 0.2 μm) of 100 images. Strong VIP and VPAC2 immunoreactivity were found in SCN (A) with VIP fibers projecting toward both the rostral part of PVN (B) and mid part of PVN (C) . VPAC1 immunoreactivity was found in both the rostral (E) and mid (F) part of PVN using ENVISION for visualization. Colocalization module (see “Materials and Methods” section) was used to determine close apposition between VIP fibers and VPAC1. Frames (G,H) represent the white frames in (D,E) , respectively. Because of amplification with biotinylated tyramide, VPAC1 in the blood vessels becomes visible (G) . 3D reconstruction of frame (H) represented in frame I shows VIP fibers making synaptic appositions (white dots indicated by arrows) on several VPAC1 receptor expressing neurons in PVN. Determination of possible synaptic appositions between VIP and VPAC1 was made using the IMARIS colocalization module (see “Materials and Methods” for details). Scale bars: (A–E) = 100 μm, (G–H) = 25 μm, (I) = 7 μm.

Article Snippet: After 15-min fixation in 4% PFA, hybridization overnight, and washing, slides were incubated with a sheep anti-digoxigenin antibody (Roche, code: 11 214 667 001; diluted 1:2,000) followed by washing and incubation with a biotinylated donkey anti–sheep antibody (Jackson ImmunoResearch code; 713-066-147, diluted 1:800), washed followed by incubation in ABC reagent (Vectastain PK6100) biotinylated tyramide (Perkin Elmer SAT 700, diluted 1:50) and finally in streptavidin conjugated Alexa Fluor 488 (Jackson ImmunoResearch; code 016-540-084, diluted 1:500).

Techniques: Amplification, Expressing