Alexa Fluor® 647 anti-Neurofilament H, Nonphosphorylated Antibody

Pricing & Availability
Clone
SMI 32 (See other available formats)
Regulatory Status
RUO
Other Names
Neurofilament heavy polypeptide, NF-H, 200 kD neurofilament protein, neurofilament triplet H protein
Isotype
Mouse IgG1
SMI-32_A647_Neurofilament_H_NF-H_Antibody_1_IHC_060618
IHC staining of Alexa Fluor® 647 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 incubated with the primary antibody at 5 µg/mL overnight at 4°C. Nuclei were counterstained with DAPI. The image was captured with a 10X objective. Scale bar: 100 µm
  • SMI-32_A647_Neurofilament_H_NF-H_Antibody_1_IHC_060618
    IHC staining of Alexa Fluor® 647 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 incubated with the primary antibody at 5 µg/mL overnight at 4°C. Nuclei were counterstained with DAPI. The image was captured with a 10X objective. Scale bar: 100 µm
  • SMI-32_A647_Neurofilament_H_NF-H_Antibody_2_IHC_060618
    IHC staining of Alexa Fluor® 647 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 incubated with the primary antibody at 5 µg/mL overnight at 4°C. Nuclei were counterstained with DAPI. The image was captured with a 40X objective. Scale bar: 50 µm
  • SMI-32_A647_Neurofilament_H_NF-H_Antibody_3_IHC_060618
    IHC staining of Alexa Fluor® 647 anti-Neurofilament H (NF-H), Nonphosphorylated antibody (clone SMI 32) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3 (Cat. No. 927801), the tissue was incubated with the primary antibody at 5 µg/mL overnight at 4°C. Nuclei were counterstained with DAPI. The image was captured with a 40X objective. Scale bar: 50 µm
  • SMI-32_A647_Neurofilament_H_NF-H_Antibody_4_IHC_060618
    IHC staining of Alexa Fluor° 647 anti-Neurofilament H (NF-H), Nonphosphorylated antibody (clone SMI 32) on formalin-fixed paraffin-embedded human cerebellum tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3 (Cat. No. 927801), the tissue was incubated with the primary antibody at 5 µg/mL overnight at 4°C. Nuclei were counterstained with DAPI. The image was captured with a 40X objective. Scale bar: 50 µm
Compare all formats See Alexa Fluor® 647 spectral data
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801711 25 µg $129.00
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801712 100 µg $323.00
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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).

Technical data sheet

Product Details

Verified 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® 647 under optimal conditions.
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
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. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.


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

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Excitation Laser
Red Laser (633 nm)
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 literature, not verified in house4.

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_2734549 (BioLegend Cat. No. 801711)
AB_2734549 (BioLegend Cat. No. 801712)

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. J Neuroimmunol. 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 on UniProt.org
Go To Top Version: 1    Revision Date: 06/06/2018

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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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