NE1022 Sigma-AldrichPhosphoDetect™ Anti-Neurofilament H Mouse mAb (SMI-31)

Safety Data Sheet (SDS) 

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision	01-March-2021 JSW
Application	ELISA (1:1000) Frozen Sections (1:1000, see comments) Immunoblotting (1:1000) Immunocytochemistry (1:1000, see comments) Paraffin Sections (1:1000, heat pre-treatment required, see comments) Immunoprecipitation (see comments)
Application Data	Detection of phosphorylated rat neurofilament H by staining frozen sections. Sample: Rat brain. Primary antibody: PhosphoDetect™ Anti-Neurofilament-H Mouse mAb (SMI-31) (Cat. No. NE1022) (1:1000). Detection: fluorescence (green) with Hoechst 33342 counterstain.
Description	Mouse monoclonal antibody supplied as purified antibody. Recognizes the ~180-200 kDa phosphorylated neurofilament H protein.
Background	Neurofilaments are a type of intermediate filament that serve as major constituent of the cytoskeleton of axons. They are the most abundant fibrillar components of the axon and are composed of three intertwined protofibrils. The neurofilament triplet proteins (NF-L, 68/70 kDa; NF-M, 160 kDa; and NF-H, 200 kDa) are found in both the central and peripheral nervous system and are usually neuron-specific. NF-H and NF-M both require the presence of NF-L protein to co-assemble. NF-H and NF-M become highly phosphorylated after newly formed neurofilaments enter the axon. Neurofilament staining is observed in neuromas, ganglioneuromas, gangliogliomas, ganglioneuroblastomas and neuroblastomas, and have also been detected in paragangliomas and adrenal and extra-adrenal pheochromocytomas. Carcinoids, neuroendocrine carcinomas of the skin, and oat cell carcinomas of the lung are also known to express neurofilaments.
Host	Mouse
Immunogen species	Rat
Immunogen	homogenized, hypothalami from Fischer 344 rat brain
Clone	SMI-31
Isotype	IgG₁
Species	Xenopus, chicken, mammalian
Positive control	Rat brain or central nervous system cytoskeletal preparations
Form	Liquid
Formulation	PBS, 0.09% Sodium Azide
Comments	Strongly recognizes phosphorylated neurofilament H and, to a lesser extent, phosphorylated neurofilament M. By immunocytochemistry this antibody broadly stains thick and thin axons and some dendrites, such as basket cell dendrites, but not Purkinje cell dendrites. Does not generally stain nerve cell bodies or other cells and tissues except peripheral axons. May also stain neuronal cell bodies in pathological conditions. Aberrant phosphorylation of neurofilament H in cell bodies can be demonstrated in neuronal cell cultures following treatment with agents that induce stress-activated protein kinase. This antibody is reported to co-immunoprecipitate neurofilament-associated kinase (NAK 115) via interaction of the antibody with the tail domain of neurofilament H. Phosphatase treatment of tissue sections or immunoblotting samples abolishes antibody reactivity. Reactivity is unaffected by trypsin treatment of samples. Tissues and cultured cells can be fixed with a variety of paraformaldehyde- or formaldehyde-containing fixatives, including Bouin's fixative. Post-fixation in cold methanol or methanol/hydrogen peroxide facilitates access of the antibody to the epitope in frozen sections or thick tissue sections fixed in 4% paraformaldehyde and in cultured cells. For staining formalin-fixed, paraffin sections it is recommended that the de-paraffinized tissue be autoclaved in dH2O for 10 min or boiled in Tris-buffered saline, pH 9.0, in a microwave for 15 min to expose the epitope. Antibody should be titrated for optimal results in individual systems.
Storage	Avoid freeze/thaw +2°C to +8°C
Do Not Freeze	Ok to freeze
Special Instructions	Following initial thaw, aliquot and freeze (-20°C).
Toxicity	Standard Handling
References	Raina, A.K., et al. 1999. Neuroreport 10, 1355. Yang, C.C., et al. 1998. Brain 121, 1089. Giasson, B.I and Mushynski, W.E. 1996. J. Biol. Chem. 271, 30404. Mirabella, M., et al. 1996. J. Neuropath. Exp. Neurol. 55, 774. Xiao, J. and Monteiro, M.J. 1994. J. Neurosci. 14, 1820.