Alexa fluor 488 donkey anti sheep igg h l

The injured birds were anesthetized at P45 via intramuscular injection of 20% barbiturate (30 μl/g body weight) and placed in a stereotaxic head holder. Fluorogold was injected into the RA (backward: 1.1–1.4 mm; left/right: 2.2–2.3 mm; depth: 2.3–2.6 mm) of one cerebral hemisphere via a glass micropipette attached to a four-channel pressure injector (MDI, PM2000B). The birds were allowed to recover for four days and then were perfused and fixed with paraformaldehyde. The brains were then cut into 40 μm sections with a freezing microtome. The sections were incubated with anti-BDNF (Chemicon, 1513P, 1:200) primary antibody overnight at 4°C. After the sections were washed, they were incubated with an Alexa Fluor 488 Donkey Anti-Sheep IgG (H+L; Molecular Probe, A-11055, 1:40) secondary antibody for 2 h at room temperature.

Human conjunctival tissue samples were from patients with SJS/TEN undergoing ocular surface reconstruction. The controls were nearly normal conjunctival tissue samples from operated patients with conjunctivochalasis.11–14 (link)
Conjunctival tissue sections mounted on slides were fixed for 10 min at 4°C with 4% PFA/0.1M phosphate-buffered saline (PBS), incubated overnight in a moist chamber at 4°C with sheep anti-human teneurin-2 polyclonal antibody (AF4578; R&D, MN, USA) or isotype control sheep IgG (R&D), and then washed in PBS without Ca and Mg [PBS(−)]. Alexa Fluor 488 donkey anti-sheep IgG (H+L) (Molecular Probes, Eugene, OR) was applied for 1 hour at room temperature. After washing the slides, antifade mounting medium with 4′,6-diamidino-2-phenylindole was applied (Vectashield; Vector Laboratories, Burlingame, CA).