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

Overview

Replacement Information

Key Specifications Table

Species ReactivityHostAntibody Type
Ch, Ma, XnMMonoclonal Antibody

Products

Catalog NumberPackaging Qty/Pack
NE1022-100UL Glass bottle 100 ul
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
FormulationPBS, 0.09% Sodium Azide
Positive controlRat brain or central nervous system cytoskeletal preparations
Quality LevelMQ100
Applications
Key Applications Enzyme-Linked Immunosorbent Assay
Frozen Sections
Immunoblotting (Western Blotting)
Immunocytochemistry
Paraffin Sections
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 Blue Ice Only
Toxicity Standard Handling
Storage +2°C to +8°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
Global Trade Item Number
Catalog Number GTIN
NE1022-100UL 04055977209907

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.

Revision01-March-2021 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
FormulationPBS, 0.09% 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 Avoid freeze/thaw
+2°C to +8°C
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.