Two antipeptide antibodies were generated in chickens against sequences shared between the mouse (AAT58219.1), rat (NP_445831.3) and human (CAA06617.1) gene products. These antibodies were affinity-purified and then mixed in equal concentrations. These antibodies have been validated with human, mouse and rat tissues.
Doublecortin (DCX, Aves Labs, 1:1000 dilution) staining (GREEN) of neuroblastoma cells in culture. RED staining is a Golgi apparatus marker. Page Balisch (University of Arizona)
Doublecortin is a microtubule-associated protein found in neuroblasts and immature neurons, especially those undergoing embryonic migrations. It was first identified as the X-chromosome-encoded gene product mutated in the human diseases Lissencephaly (in males) and Periventricular Heterotopia (in females), diseases caused by failure of neocortical neuroblast migration. The human protein itself is 360 amino acids (40,044 daltons) and contains doublecortin motifs near its N-terminus that are believed to mediate the microtubule binding.
DCX DBCN LISX
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 two synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the doublecortin gene product, but are shared between the human (CAA06617.1, NCBI) and mouse (AAT58219.1, NCBI) sequences. After repeated injections, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column, and the concentrations of the eluates adjusted to 200 µg/mL. Finally, equal volumes of these affinity-purified antipeptide antibodies were mixed, and the preparation was filter-sterilized.
This anti-peptide antibody mixture was analyzed by 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.
Neuronal migration protein doublecortin (Doublin) (Lissencephalin-X) (Lis-X)
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