| Description |  | Purified form of monoclonal antibody to neurofilaments, phosphorylated epitope, Clone SMI 31 |
| Intended Use | | **Research Use Only (RUO)** This product is sold for laboratory research use only, not for human or in-vivo use. |
| Clone | | SMI-31 |
| Form | | Purified Antibody (in PBS + Thimerosal) |
| Host | | Mouse |
| Species Reactivity | | Neurofilaments, Phosphorylated (SMI 31) Monoclonal Antibody, Purified Mammalian, Chicken, Xenopus |
| IsoType | | IgG1 |
| [Ab] | | 1 mg/mL |
| Specificity | | SMI 31 reacts with a phosphorylated epitope in extensively phosphorylated neurofilament H and, to a lesser extent, with neurofilament M in most mammalian species, as well as in chicken and frog (Xenopus). Immunocytochemically, SMI 31 reacts broadly with thick and thin axons and some dendrites such as basket cell dendrites, but not Purkinje cell dendrites. Nerve cell bodies are generally unreactive. Other cells and tissues are unreactive except for peripheral axons. Phosphatase treatment of tissue sections or Western blots abolishes reaction with SMI 31. Staining is unaffected by trypsin. In pathological conditions, reaction with SMI 31 may be found also in neuronal cell bodies. Aberrant phosphorylation of neurofilament H in cell bodies can be demonstrated in neuronal cell cultures with SMI 31 by agents that induce stress-activated protein kinase. In its reaction with paired helical filaments in hereditary inclusion body myopathy, SMI 31 colocalizes with nitric oxide synthase, suggesting that oxidative stress may play a role in the pathogenic cascade of such degenerative diseases. SMI 31 co-immunoprecipitates neurofilament-associated kinase (NAK 115) via reaction of the antibody with the tail domain of neurofilament H. |
| 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 31 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 31 reacts on Western blots, paraffin, vibratome and frozen tissue sections as well as with neuronal cell cultures. Tissues and cultures can be fixed in a variety of paraformaldehyde or formaldehyde-containing fixatives including Bouins. "Recovery" of antigen in tissue that has been formalin-fixed for long periods of time may be accomplished by autoclaving of deparaffinized sections 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). Post-fixation in cold methanol or methanol/hydrogen peroxide greatly facilitates access of SMI 31 in frozen sections or thick sections of tissue perfused with 4% paraformaldehyde and in tissue cultures. These antibodies can be used for neurofilament detection in trypsin-treated tissue. |
| Storage | | Neurofilaments, Phosphorylated (SMI 31) Monoclonal Antibody, Purified 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 | | Raina AK, Takeda A, Nunomura A, Perry G, Smith MA. Genetic evidence for oxidative stress in Alzheimers disease. Neuroreport 10:1, 1999. Yang CC, Alvarez RR, Engel WK, Heller SL, Askansas. Nitric oxide-induced oxidative stress in autosomal recessive and dominant inclusion-body myopathies. Brain 121:1089, 1998. Giasson BI, Mushynski WE. Aberrant stress-induced phosphorylation of perikaryal neurofilaments. J Biol Chem 271:30404, 1996. Mirabella M, Alvarez RB, Bilak M, Engel WK, Askansas V. Differences in expression of phosphorylated tau epitopes between sporadic inclusion-body myositis and hereditary inclusion-body myopathies. J Neurpoath Exp Neurol 55:774, 1996. Xiao J, Monteiro MJ. Identification and characterization of a novel (115 kDa) neurofilament-associated kinase. J Neurosci 14:1820, 1994. |
| Warranty/Conditions | | Covance products may not be resold or modified for resale without prior written approval. |
