An antipeptide antibody was generated in chickens against a sequence shared between the mouse (O09118) and human (O95631) gene products. Antibodies were affinity-purified and the concentration adjusted to 100 µg/mL. These antibodies have been validated with human, mouse and rat tissues.
Neurosphere (organoid) cultures of e13 mouse brain were cultured for 2 weeks, and then paraformaldehyde (2%) fixed. After extensive washing, the cultures were incubated with Netrin-1 antibody (Cat.# NET; 1:500 dilution), washed, and then treated with fluorescein-labeled goat anti-chicken IgY (Aves Cat.# F-1005; 1:500 dilution). Hoda Ilias, Aves Labs.
Murine netrin-1 precursor is a 67,768 dalton protein (604 amino acids) expressed by subpopulations of cells within the developing CNS, as well as in various non-neural tissues, including the gut, liver, heart, and prostate. In the embryonic CNS, netrin-1 is expressed by cells of the floor plate, and acts to attract axons of "commissural neurons" from the dorsal horn gray matter. Netrin-1 is also believed to play a role in the growth of motor axons in the PNS.
Human, Mouse, Rat
Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze these antibodies unless you want to store them for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half of its binding activity is lost.
Chickens were immunized with a synthetic peptide-keyhole limpet hemocyanin (KLH) conjugate. This synthetic peptide corresponded to the netrin-1 gene product, but was shared between the human (O_095631, NCBI) and mouse (O_09118, NCBI) sequences. After repeated injections into the hens, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column, the concentrations of the eluate adjusted to 100 µg/mL, and the preparation filter-sterilized.
WB: Not Recommended
Quality assurance analysis was performed using immunohistochemistry (at a dilution of 1:2000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.
These antibodies are to be used as research laboratory reagents and are not for use as diagnostic or therapeutic reagents in humans.
Netrin-1 (Epididymis tissue protein Li 131P)
UniProt (Immunogen Species): O95631
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