Preparation And Storage
Recommended Assay Procedures
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Multipotential stem cells in the neural tube of the developing embryo give rise to the different neuronal cell types of the brain. Nestin is an intermediate filament protein which is abundantly expressed in neuroepithelial stem cells early in embryogenesis, but is absent from nearly all mature central nervous system (CNS) cells. After its down-regulation, GFAP and neurofilaments are expressed in differentiated astrocytes and neurons, respectively. Initial studies showed that the mouse anti-rat nestin antibody (clone Rat 401) recognized transient radial glial cells and dividing neuroepithelial stem cells in the embryonic rat CNS. Reportedly, Rat 401 has been used to analyze nestin expression in the developing rat nervous system and in immortalized CNS precursor cell lines (e.g such as in E11 rat CNS stem cells, but lost by postnatal day 6 (P6) in spinal cord and by P21 in the cerebellum). Although not expressed in normal adult CNS cells, nestin has been reportedly to be detectable in a variety of CNS tumors, suggesting that these tumors share gene expression patterns with primitive, undifferentiated CNS cells. Rat 401 positive cells have been reported to be found throughout the developing neural tube, but not in the adult CNS. More than 90% of dissociated E11 neural tube cells have been reported to be Rat 401 positive. Rat 401 recognizes rat nestin and has been reported not to cross-react with human nestin. Due to differential tissue expression, this antibody may recognize nestin as a doublet within a range of 198-260 kD.
Development References (9)
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Dahlstrand J, Collins VP, Lendahl U. Expression of the class VI intermediate filament nestin in human central nervous system tumors. Cell. 1992; 52(19):5334-5341. (Biology). View Reference
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Dahlstrand J, Zimmerman LB, McKay RD, Lendahl U. Characterization of the human nestin gene reveals a close evolutionary relationship to neurofilaments. J Cell Sci. 1992; 103(2):589-597. (Biology). View Reference
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Frederiksen K, McKay RD. Proliferation and differentiation of rat neuroepithelial precursor cells in vivo. J Neurosci. 1988; 8(4):1144-1151. (Biology: Flow cytometry, Fluorescence microscopy). View Reference
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Geschwind DH, Hockfield S. Identification of proteins that are developmentally regulated during early cerebral corticogenesis in the rat. J Neurosci. 1989; 9(12):4303-4317. (Biology: Immunohistochemistry, Western blot). View Reference
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Hockfield S, McKay RD. Identification of major cell classes in the developing mammalian nervous system. J Neurosci. 1985; 5(12):3310-3328. (Immunogen: Electron microscopy, Immunohistochemistry). View Reference
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Lendahl U, Zimmerman LB, McKay RD. CNS stem cells express a new class of intermediate filament protein. Cell. 1990; 60(4):585-595. (Biology: Fluorescence microscopy). View Reference
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Renfranz PJ, Cunningham MG, McKay RD. Region-specific differentiation of the hippocampal stem cell line HiB5 upon implantation into the developing mammalian brain. Cell. 1991; 66(4):713-729. (Biology: Fluorescence microscopy). View Reference
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Tohyama T, Lee VM, Rorke LB, Marvin M, McKay RD, Trojanowski JQ. Monoclonal antibodies to a rat nestin fusion protein recognize a 220-kDa polypeptide in subsets of fetal and adult human central nervous system neurons and in primitive neuroectodermal tumor cells. Am J Pathol. 1993; 143(1):258-268. (Biology). View Reference
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Tohyama T, Lee VM, Rorke LB, Marvin M, McKay RD, Trojanowski JQ. Nestin expression in embryonic human neuroepithelium and in human neuroepithelial tumor cells. Lab Invest. 1992; 66(3):303-313. (Biology). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.
Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
