Alexa Fluor® 594 anti-Neurofilament H (NF-H), Nonphosphorylated Antibody

Pricing & Availability
Clone
SMI 32 (See other available formats)
Other Names
Neurofilament heavy polypeptide, NF-H, 200 kD neurofilament protein, neurofilament triplet H protein
Isotype
Mouse IgG1, κ
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Product Citations
publications
SMI-32_A594_NeuroH_Antibody_11_122817
IHC staining of Alexa Fluor® 594 anti-Neurofilament H (NF-H), Nonphosphorylated antibody (clone SMI 32) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3 (Cat. No. 927801), the tissue was blocked with a buffer containing 5% Normal Goat Serum, 1% BSA and 0.25% Triton-X-100 in PBS. The tissue was then incubated with 5 µg/ml of the primary antibody overnight at 4°C. Nuclei were counterstained with DAPI. The image was captured with a 40x objective. Scale bar: 50 µm.
  • SMI-32_A594_NeuroH_Antibody_11_122817
    IHC staining of Alexa Fluor® 594 anti-Neurofilament H (NF-H), Nonphosphorylated antibody (clone SMI 32) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3 (Cat. No. 927801), the tissue was blocked with a buffer containing 5% Normal Goat Serum, 1% BSA and 0.25% Triton-X-100 in PBS. The tissue was then incubated with 5 µg/ml of the primary antibody overnight at 4°C. Nuclei were counterstained with DAPI. The image was captured with a 40x objective. Scale bar: 50 µm.
  • SMI-32_A594_NeuroH_Antibody_22_122817
    IHC staining of Alexa Fluor® 594 anti-Neurofilament H (NF-H), Nonphosphorylated antibody (clone SMI 32) on frozen mouse brain tissue. The tissue was permeabilized with 0.25% Triton-X-100 in PBS, then blocked with a buffer containing 5% Normal Goat Serum, 1% BSA and 0.25% Triton-X-100 in PBS. The tissue was then incubated with 5 µg/ml of the primary antibody overnight at 4°C. Nuclei were counterstained with DAPI. The image was captured with a 40x objective. Scale bar: 50 µm
  • SMI-32_A594_NeuroH_Antibody_33_122817
    ICC staining of Alexa Fluor® 594 anti-Neurofilament H (NF-H), Nonphosphorylated antibody (clone SMI 32) on SH-SY5Y neuroblastoma cells. The cells were fixed with 100% Methanol, permeabilized with 0.1% Triton-X-100 and 0.1% BSA in PBS, and blocked with 2% Normal Goat Serum and 0.02% BSA in PBS. The cells were then incubated with 5 µg/mL of the primary antibody overnight at 4°C. Nuclei were counterstained with DAPI. The image was captured with a 40x objective. Scale bar: 50 µm.
See Alexa Fluor® 594 spectral data
Cat # Size Price Quantity Avail. Save
801709 25 µg $110
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801710 100 µg $275
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Description

Neurofilaments (NF) 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 NF (NF-L) runs at 68-70 kD. 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. 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

Product Details

Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 under optimal conditions. The solution is free of unconjugated Alexa Fluor® 594.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

IHC-P - Quality tested
ICC, IHC-F - Validated

Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry (formalin-fixed paraffin-embedded), a concentration range of 5.0 - 10.0 µg/ml is suggested. For immunocytochemistry, a concentration range of 5.0 - 10.0 μg/ml is recommended. For immunohistochemical staining on frozen tissue sections, a concentration range of 5.0 - 10.0 µg/ml is suggested.It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Application Notes

Additional reported applications (for the relevant formats) include Western blotting6, immunohistochemistry4,5, immunocytochemistry1,2,3, 7, array tomography8.

Cross-reactivity to monkey tissue has been reported in the literature4.

This antibody reacts with a nonphosphorylated epitope in neurofilament H of most mammalian species. The reaction is masked when the epitope is phosphorylated. The staining of isolated neurofilament preparations is greatly intensified upon dephosphorylation. Immunocytochemically, SMI 32 visualizes neuronal cell bodies, dendrites, and some thick axons in the central and peripheral nervous systems. However, thin axons are not revealed. Other cells and tissues are unreactive. The antibody distinguishes three subdivisions of the macaque precentral motor cortex. The greater size of the left versus the right superior temporal lobe was found to be due to increased axonal myelination and not due to increased number of glial cells or SMI 32-enumerated neurons, suggesting that the specialization for language in the left temporal lobe is related to increased speed of signal transmission. In cultures of murine cortex, SMI 32 labels a neuronal population with enhanced vulnerability to kainate toxicity most of which are GABAergic and reveal kainate-activated Ca2+ uptake.

Application References

(PubMed link indicates BioLegend citation)
  1. Chang Q, Martin LJ. 2011. J. Neurosci., 31:2815-27. (ICC) PubMed
  2. Stevens HE, et al. 2010. J. Neurosci. 30:5590-602. (ICC) PubMed
  3. Kiryu-Seo S, et al. 2010. J. Neurosci. 30:6658-66. (ICC) PubMed
  4. Redondo J, et al. 2015. Brain Pathol. 25(6):692. (IHC-P) PubMed
  5. Feng L, et al. 2017. eNeuro. 4(1): 0331-16.2016. (IHC-P) PubMed
  6. Feng L, et al. 2014. Invest Ophthalmol Vis Sci. 54(2): 1106:1117. (IHC-P) PubMed
  7. Theotokis, et al. 2016. J. Neuroinflammation 13(1):265 (IHC-P)
  8. Bennett, et al. 2015. J. Neurosci. Methods 245:25-36 (Array Tomography)
  9. Petzold A, et al. 2011. Brain 134:464. (WB) PubMed
RRID
AB_2721392 (BioLegend Cat. No. 801709)
AB_2721393 (BioLegend Cat. No. 801710)

Antigen Details

Structure
Neurofilament H has an apparent molecular mass of 200-220 kD.
Distribution

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

Function
NF-H Neurofilaments are the major components of the neuronal cytoskeleton. They provide axonal support and regulate axon diameter. Phosphorylation of NF-H results in the formation of interfilament cross bridges that are important in the maintenance of axonal caliber.
Ligand Receptor
Phosphorylation seems to play a major role in the functioning of the larger neurofilament polypeptides (NF-M and NF-H), the levels of phosphorylation result in changes to the neurofilament function.
Cell Type
Mature Neurons
Biology Area
Cell Biology, Neuroscience, Neuroscience Cell Markers
Molecular Family
Intermediate Filaments, Phospho-Proteins
Antigen References
  1. Turner M, et al. 2015. Journal of Neuroimmunology. 285: 4. PubMed
  2. Pagliarini V, et al. 2015. J. Cell Biol.. 211: 77. PubMed
  3. Petzold A, et al. 2011. Brain 134. (WB) PubMed 
  4. Yuan A, et al. 2016. Brain Res Bull  126(3): 334.
  5. Parlakian A, et al. 2016. Rev Neurol. 172(10): 607.
  6. Li D, et al. 2016. Front Aging Neurosci. 8: 290.
  7. Costa J, et al. 2016. Clin Chim Acta. 455: 7.
  8. Lad SP, et al. 2010.  J Stroke Cerebrovasc Dis. 21(1): 30.
Gene ID
4744 View all products for this Gene ID
UniProt
View information about Neurofilament H (NF-H) on UniProt.org

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