HRP anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated Antibody

Pricing & Availability
Clone
SMI 35 (See other available formats)
Other Names
Neurofilament heavy polypeptide, NF-H, 200 kD neurofilament protein, neurofilament triplet H protein
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
SMI-35_HRP_Neurofilament_Phosphorylated_Antibody_IHC_062017
IHC staining of HRP anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated Antibody (clone SMI 35) on formalin-fixed paraffin-embedded human cerebellum brain tissue. After antigen retrieval using Retrieve-All Antigen Unmasking System 3, the tissue was incubated with the primary antibody at 5 µg/mL for one hour at room temperature. DAB was used for detection followed by hematoxylin counterstaining and Bluing solution counterstaining.
  • SMI-35_HRP_Neurofilament_Phosphorylated_Antibody_IHC_062017
    IHC staining of HRP anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated Antibody (clone SMI 35) on formalin-fixed paraffin-embedded human cerebellum brain tissue. After antigen retrieval using Retrieve-All Antigen Unmasking System 3, the tissue was incubated with the primary antibody at 5 µg/mL for one hour at room temperature. DAB was used for detection followed by hematoxylin counterstaining and Bluing solution counterstaining.
  • SMI-35_HRP_Neurofilament_Phosphorylated_Antibody_WB_062017
    Western blot of HRP anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated (clone SMI 35), and Isotype-matched IgG1 control. Lane 1: Molecular weight marker; Lane 2: 20 µg of human brain lysate; Lane 3: 20µg of rat brain lysate; Lane 4: 20µg of mouse brain lysate. Blots were incubated with 5.0µg/mL of each primary antibody overnight at 4°C. The isotype-matched control blot was further incubated with horseradish peroxidase labeled goat anti-mouse secondary antibody. Enhanced chemiluminescence was used as the detection system.
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835607 25 µg 168€
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835608 100 µg 424€
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Description

Neurofilaments (NFs) are approximately 10 nanometer intermediate filaments found in neurons. They are a major component of the neuronal cytoskeleton, and function primarily to provide structural support for the axon and to regulate the axon diameter. There are three major NF subunits, and the names given to these subunits are based upon the apparent molecular mass of the mammalian subunits on SDS-PAGE. The light or lowest neurofilament (NF-L) runs at 68-70 kD. The medium or middle (NF-M) runs at about 145-160 kD, and the heavy or highest (NF-H) runs at 200-220 kD. However, the actual molecular weight of these proteins is considerably lower due to the highly charged C-terminal regions of the molecules. The level of NF gene expression correlates with the axonal diameter, which controls how fast electrical signals travel down the axon. Mutant mice with NF abnormalities have phenotypes resembling amyotrophic lateral sclerosis. NF immunostaining is common in diagnostic neuropathology. It is useful for differentiating neurons (positive for NF) from the glia (negative for NF).

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
This antibody is provided in 50% glycerol in aqueous buffered solution with preservatives.
Preparation
The antibody was purified by affinity chromatography and conjugated with HRP under optimal conditions.
Concentration
0.5 mg/ml
Storage & Handling
Upon receipt, the antibody solution should be stored undiluted at -20°C, and protected from prolonged exposure to light.
Application

IHC-P - Quality tested
WB - Validated

Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 5.0 - 10 µg/ml is suggested. For Western blotting, the suggested use of this reagent is 1.0 - 5.0 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Additional reported applications (for the relevant formats of this clone): include ELISA Capture1-3, immunohistochemical staining on frozen tissue sections and immunofluorescence staining.

Clone SMI 35 reacts with highly phosphorylated neurofilaments, as well as with low degrees of phosphorylation. It primarily reacts with neurofilament H and with neurofilament M to a lesser extent.

Notes: On two dimensional gels, this antibody detects a band extending from the phosphorylated neurofilament position at 200 kD (pI 5.1) toward the non-phosphorylated position at 170 kD (pI 6.2).

Application References

(PubMed link indicates BioLegend citation)
  1. Petzold A. 2013. J. Neuroimmunol. 262:(1-10). (ELISA)
  2. Lu CH, et al. 2012. PLoS One. 7:e40998. (ELISA)
  3. Steinacker P, et al. 2011. PLoS One. 8:e23600. (ELISA)
  4. Poltorak M, et al. 1993. J. Neurosci. 13:2217. (IHC-F)
  5. Petzold A, et al. 2011. Brain. 134:464. (WB) PubMed
RRID
AB_2686948 (BioLegend Cat. No. 835607)
AB_2686949 (BioLegend Cat. No. 835608)

Antigen Details

Structure
The medium or middle NF (NF-M) runs at about 145-160 kD, and the heavy or highest NF (NF-H) runs at 200-220 kD.
Distribution

Tissue Distribution: CNS, peripheral nerves and glandular cells of the prostate
Cellular Distribution: Cytoskeleton, nucleus, cytosol, and mitochondrion

Function
Neurofilaments are the major components of the neuronal cytoskeleton. They provide axonal support and regulate axon diameter.
Interaction
Cell bodies and dendrites are generally unstained. Other cells and tissues are unreactive except for peripheral axons.
Cell Type
Mature Neurons
Biology Area
Cell Biology, Cell Motility/Cytoskeleton/Structure, Neuroscience, Neuroscience Cell Markers
Molecular Family
Intermediate Filaments, Phospho-Proteins
Antigen References
  1. Petzold A. 2012. Mult Scler Int. 2012:217802. PubMed
  2. Gresle MM, et al. 2011. Mult Scler Int. 2011: 315406. PubMed
Gene ID
4744 View all products for this Gene ID
UniProt
View information about Neurofilament H & M (NF-H/NF-M), Phospho on UniProt.org

Related FAQs

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Go To Top Version: 2    Revision Date: 01/29/2018

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