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.
Volume: 1000 µL
Concentration: 200 µg/mL
Clonality: Polyclonal
Host Species: Chicken
Species Reactivity: Human, Mouse, Rat
Applications: ICC, IHC
Protein Name / Synonyms: Neuronal migration protein doublecortin (Doublin) (Lissencephalin-X) (Lis-X)
Target Description: 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.
Gene ID: DCX DBCN LISX
Antibody Registry ID (RRID): AB_2313540
Physical State: Liquid
Production Notes: 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.
Validation and Application Notes
IHC Dilution Range: 1:1000-1:2000
Quality Control
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.
Storage
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.
NOTE
Aves Labs products are intended for use as research laboratory reagents. They are not intended for use as diagnostic or therapeutic reagents in humans.
Citations
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- Mehta SR, Tom CM, Wang Y, Bresee C, Rushton D, Mathkar PP, Tang J, Mattis VB (2018), 'Human Huntington’s Disease iPSC-Derived Cortical Neurons Display Altered Transcriptomics, Morphology, and Maturation.' Cell Reports. 10.1016/j.celrep.2018.09.076.
- A Alachkar, L Wang, R Yoshimura, AR Hamzeh, Z Wang, N Sanathara, SM Lee, X Xu, GW Abbott, and O Civelli (2017), 'Prenatal one-carbon metabolism dysregulation programs schizophrenia-like deficits.' Molecular Psychiatry. 10.1038/mp.2017.164.