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

This PhosphoDetect™ Anti-Neurofilament H Mouse mAb (SMI-31) is validated for use in ELISA, Frozen Sections, Immunoblotting, ICC, Paraffin Sections, IP for the detection of Neurofilament H.

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NE1022
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Overview

Key Spec Table

Species ReactivityHostAntibody Type
Ch, Ma, XnMMonoclonal Antibody

Products

Catalogue NumberPackaging Qty/Pack
NE1022-100UL Glass bottle 100 ul
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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
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
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).
Global Trade Item Number
Catalogue Number GTIN
NE1022-100UL 04055977209907