| Description |  | Purified Monoclonal Antibody to Non-Phosphorylated Neurofilaments |
| Intended Use | | **Research Use Only (RUO)** This product is sold for laboratory research use only, not for human or in-vivo use. |
| Clone | | SMI-32 |
| Form | | Purified Antibody |
| Host | | Mouse |
| Species Reactivity | | Mammalian |
| IsoType | | Neurofilament H Non-Phosphorylated (SMI 32) Monoclonal Antibody, Purified IgG1 |
| Specificity | | SMI 32 reacts with a nonphosphorylated epitope in neurofilament H of most mammalian species. The reaction is masked when the epitope is phosphorylated. Staining of isolated neurofilament preparations is greatly intensified upon dephosphorylation. Immunocytochemically, SMI 32 visualizes neuronal cell bodies, dendrites and some thick axons in the central and peripheral nervous systems, but thin axons are not revealed. Other cells and tissues are unreactive. The antibody distinguishes three subdivisions of the macaque precentral motor cortex. The greater size of the left versus the right superior temporal lobe was found to be due to increased axonal myelination and not due to increased number of glial cells or SMI 32-enumerated neurons, suggesting that the specialization for language in the left temporal lobe is related to increased speed of signal transmission. In cultures of murine cortex, SMI 32 labels a neuronal population with enhanced vulnerability to kainate toxicity most of which are GABAergic and reveal kainate-activated Ca2+ uptake. |
| Uses | | This antibody is effective in immunoblotting, immunohistochemistry (IHC), immunocytochemistry and ELISA. |
| Suggested Working Dilution | | The optimal working dilution should be determined for each specific assay condition.
The extent of permissible dilution of SMI 32 beyond those recommended for general application depends upon nature and concentration of the antigen examined, species of the antigen, method of fixation and kind of section examined. Tissue Preparation: SMI 32 reacts on Western blots, paraffin, vibratome and frozen tissue sections as well as cell cultures. Tissues and cultures can be fixed in a variety of paraformaldehyde or formaldehyde-containing fixatives such as Bouins. Reaction is poor in material fixed in glutaraldehyde/paraformaldehyde. In formalin-fixed, paraffin-embedded tissue, reaction with SMI 32 can be enhanced by autoclaving deparaffinized sections for 10 min in distilled water (Shin et al, Lab Invest, 64:693, 1991) or by boiling sections or tissue blocks immersed in tris buffered saline, pH 9.0, in a microwave oven for 15 min (Evers and Uylings, J Neurosci Methods 72:197, 1997). Trypsin pretreatment abolishes reactivity with these antibodies. Post-fixation in cold methanol or methanol/hydrogen peroxide facilitates access of SMI 32 to neurons in frozen sections or thick sections of tissue perfused with 4% paraformaldehyde and in tissue cultures. |
| Notes | | Neurofilament H Non-Phosphorylated (SMI 32) Monoclonal Antibody, Purified SMI 32 visualizes two bands (200 and 180 kDa) which merge into a single NFH line on two-dimensional blots. |
| Storage | | Store at -20°C. Upon initial thawing, apportion into working aliquots and store at -20°C. Avoid repeated freeze-thaw cycles to prevent denaturing the antibody. For long-term storage, keep the antibody at -80°C. |
| References | | Trapp BD, Peterson J, Ransohoff RM, Rudick R, Mörk S, Bö L. Axonal transection in the lesions of multiple sclerosis. N Eng J Med 338:278-85, 1998. King CE, Jacobs I, Dickson TC, Vickers JC. Physical damage to rat cortical axons mimics early Alzheimers neuronal pathology. Neuroreport 8:1663-5, 1997. Campbell MJ, Hof PR, Morrison JH. A subpopulation of primate cortical neurons is distinguished by somatodendritic distribution of neurofilament protein. Brain Res 539:133, 1991. Campbell MJ, Morrison J. Monoclonal antibody to neurofilament protein (SMI 32) labels a subpopulation of pyramidal axons in human and monkey neocortex. J Comp Neurol 282:191, 1990. Sternberger LA, Sternberger NH. Monoclonal antibodies distinguish phosphorylated and non-phosphorylated forms of neurofilaments in situ. PNAS USA 80:6126-30, 1983. |
| Warranty/Conditions | | Covance products may not be resold or modified for resale without prior written approval. |
