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.

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Citations

• Smagin DA, Park JH, Michurina TV, Peunova N, Glass Z, Sayed K, Bondar NP, Kovalenko IN, Kudryavtseva NN, Enikolopov G (2015), 'Altered Hippocampal Neurogenesis and Amygdalar Neuronal Activity in Adult Mice with Repeated Experience of Aggression.' Frontiers in Neurosciene. 10.3389/fnins.2015.00443.
• Lim RG, Salazar LL, Wilton DK, King AR, Stocksdale JT, Sharifabad D, Lau AL, Stevens B, Reidling JC, Winokur ST, Casale MS, Thompson LM, Pardo M, Díaz-Barriga AGG, Straccia M, Sanders P, Alberch J, Canals JM, Kaye JA, Dunlap M, Jo L, May H, Mount E, Anderson-Bergman C, Haston K, Finkbeiner S, Kedaigle AJ, Gipson TA, Yildirim F, Ng CW, Milani P, Housman DE, Fraenkel E, Allen ND, Kemp PJ, Atwal RS, Biagioli M, Gusella JF, MacDonald ME, Akimov SS, Arbez N, Stewart J, Ross CA, Mattis VB, Tom CM, Ornelas L, Sahabian A, Lenaeus L, Mandefro B, Sareen D, Svendsen CN (2017), 'Developmental alterations in Huntington's disease neural cells and pharmacological rescue in cells and mice.' Nature Neuroscience. 10.1038/nn.4532.
• Mellios N, Feldman DA, Sheridan SD, Ip JPK, Kwok S, Amoah SK, Rosen B, Rodriguez BA, Crawford B, Swaminathan R, Chou S, Li Y, Ziats M, Ernst C, Jaenisch R, Haggarty SJ, Sur M (2017), 'MeCP2-regulated miRNAs control early human neurogenesis through differential effects on ERK and AKT signaling.' Molecular Psychiatry. 10.1038/mp.2017.86.
• 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.

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