559844 Anti-Neurofilament H Non-Phosphorylated Mouse mAb (SMI-32)

This Anti-Neurofilament H Non-Phosphorylated Mouse mAb (SMI-32) is validated for use in ELISA, Frozen Sections, WB, ICC, Paraffin Sections for the detection of Neurofilament H Non-Phosphorylated.

559844
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      Description
      OverviewRecognizes the non-phosphorylated ~180 kDa-200 kDa neurofilament H protein in rat central nervous system (CNS) cytoskeletal preparations.
      Catalogue Number559844
      Brand Family Calbiochem®
      References
      ReferencesTrapp, B.D., et al. 1998. N. Engl. J. Med. 338, 278.
      King, C.E., et al. 1997. Neuroreport. 8, 1663.
      Campbell, M.J., et al. 1991. Brain Res. 539, 133.
      Campbell, M.J., et al. 1989. J. Comp. Neurol. 282, 191.
      Sternberger, L.A., et al. 1983. Proc. Natl. Acad. Sci. USA 80, 6126.
      Product Information
      FormLiquid
      FormulationIn PBS
      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, see comments)
      Immunocytochemistry (1:1000, see comments)
      Paraffin Sections (1:1000, heat pre-treatment required, see comments)
      Biological Information
      Immunogenhomogenized hypothalami from Fischer 344 rat brain
      ImmunogenRat
      CloneSMI-32
      HostMouse
      IsotypeIgG₁
      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 +2°C to +8°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Special InstructionsOnly recognizes non-phosphorylated neurofilament H. By immunocytochemistry this antibody stains neuronal cell bodies, dendrites, and some thick axons in the central and peripheral nervous systems, but does not stain thin axons or other cells and tissues. Tissues and cultured cells can be fixed in a variety of paraformaldehyde- or formaldehye-containing fixatives, such as Bouin's fixative. Antibody reactivity is poor in glutaraldehyde/paraformaldehyde-fixed samples. For staining formalin-fixed, paraffin sections it is recommended that 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. Trypsin pre-treatment abolishes antibody binding to the epitope. Post-fixation in cold methanol or methanol/hydrogen peroxide facilitates access of the antibody to the neurons in frozen sections or thick sections fixed in 4% paraformaldehyde and in cultured cells. By immunoblotting this antibody detects two bands, ~180 and ~200 kDa, which merge into a single NFH line on two-dimensional gels. Antibody should be titrated for optimal results in individual systems.
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      Supplemental Information
      Specifications

      Documentation

      Anti-Neurofilament H Non-Phosphorylated Mouse mAb (SMI-32) SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overview
      Trapp, B.D., et al. 1998. N. Engl. J. Med. 338, 278.
      King, C.E., et al. 1997. Neuroreport. 8, 1663.
      Campbell, M.J., et al. 1991. Brain Res. 539, 133.
      Campbell, M.J., et al. 1989. J. Comp. Neurol. 282, 191.
      Sternberger, L.A., et al. 1983. Proc. Natl. Acad. Sci. USA 80, 6126.
      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.

      Revision09-May-2018 JSW
      ApplicationELISA (1:1000)
      Frozen Sections (1:1000, see comments)
      Immunoblotting (1:1000, see comments)
      Immunocytochemistry (1:1000, see comments)
      Paraffin Sections (1:1000, heat pre-treatment required, see comments)
      DescriptionMouse monoclonal antibody supplied as affinity purified antibody. Recognizes the ~180-200 kDa non-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-32
      IsotypeIgG₁
      Speciesmammalian
      FormLiquid
      FormulationIn PBS
      Storage +2°C to +8°C
      Avoid freeze/thaw
      Do Not Freeze Ok to freeze
      Special InstructionsOnly recognizes non-phosphorylated neurofilament H. By immunocytochemistry this antibody stains neuronal cell bodies, dendrites, and some thick axons in the central and peripheral nervous systems, but does not stain thin axons or other cells and tissues. Tissues and cultured cells can be fixed in a variety of paraformaldehyde- or formaldehye-containing fixatives, such as Bouin's fixative. Antibody reactivity is poor in glutaraldehyde/paraformaldehyde-fixed samples. For staining formalin-fixed, paraffin sections it is recommended that 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. Trypsin pre-treatment abolishes antibody binding to the epitope. Post-fixation in cold methanol or methanol/hydrogen peroxide facilitates access of the antibody to the neurons in frozen sections or thick sections fixed in 4% paraformaldehyde and in cultured cells. By immunoblotting this antibody detects two bands, ~180 and ~200 kDa, which merge into a single NFH line on two-dimensional gels. Antibody should be titrated for optimal results in individual systems.
      Toxicity Standard Handling
      ReferencesTrapp, B.D., et al. 1998. N. Engl. J. Med. 338, 278.
      King, C.E., et al. 1997. Neuroreport. 8, 1663.
      Campbell, M.J., et al. 1991. Brain Res. 539, 133.
      Campbell, M.J., et al. 1989. J. Comp. Neurol. 282, 191.
      Sternberger, L.A., et al. 1983. Proc. Natl. Acad. Sci. USA 80, 6126.

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