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

Overview

Replacement Information

Key Specifications Table

Species ReactivityHostAntibody Type
Ch, Ma, Xn M Monoclonal Antibody

Pricing & Availability

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NE1022-100UL
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      Description
      OverviewRecognizes the ~180-200 kDa phosphorylated neurofilament H protein in rat central nervous system (CNS) cytoskeletal preparations. Also weakly recognizes phosphorylated NF-M protein.
      Catalogue NumberNE1022
      Brand Family Calbiochem®
      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.
      References
      ReferencesRaina, 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.
      Product Information
      FormLiquid
      FormulationIn PBS and Thimerosal
      Positive controlRat brain or central nervous system cytoskeletal preparations
      Preservative≤0.1% sodium azide
      Quality LevelMQ100
      Applications
      Key Applications Immunocytochemistry
      Paraffin Sections
      Enzyme-Linked Immunosorbent Assay
      Frozen Sections
      Immunoblotting (Western Blotting)
      Application NotesELISA (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 CommentsStrongly 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.
      Biological Information
      Immunogenhomogenized, hypothalami from Fischer 344 rat brain
      ImmunogenRat
      CloneSMI-31
      HostMouse
      IsotypeIgG₁
      Species Reactivity
      • Chicken
      • Mammals
      • Xenopus
      Antibody TypeMonoclonal Antibody
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Dry Ice Only
      Toxicity Standard Handling
      Storage -20°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      PhosphoDetect™ Anti-Neurofilament H Mouse mAb (SMI-31) SDS

      Title

      Safety Data Sheet (SDS) 

      PhosphoDetect™ Anti-Neurofilament H Mouse mAb (SMI-31) Certificates of Analysis

      TitleLot Number
      NE1022

      References

      Reference overview
      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.
      Data Sheet

      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.

      Revision11-October-2016 JSW
      ApplicationELISA (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.
      DescriptionMouse monoclonal antibody supplied as purified antibody. Recognizes the ~180-200 kDa phosphorylated neurofilament H protein.
      BackgroundNeurofilaments 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.
      HostMouse
      Immunogen speciesRat
      Immunogenhomogenized, hypothalami from Fischer 344 rat brain
      CloneSMI-31
      IsotypeIgG₁
      SpeciesXenopus, chicken, mammalian
      Positive controlRat brain or central nervous system cytoskeletal preparations
      FormLiquid
      FormulationIn PBS and Thimerosal
      Preservative≤0.1% sodium azide
      CommentsStrongly 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 -20°C
      Avoid freeze/thaw
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Toxicity Standard Handling
      ReferencesRaina, 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.