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Purified anti-Neurofilament H & M (NF-H/NF-M), Phosphorylated Antibody

Pricing & Availability
Clone
SMI 310 (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, κ
Ave. Rating
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Product Citations
publications
SMI-310_PURE_Neurofilament_Phosphorylated_Antibody_1_123118_updated.png
IHC staining of purified anti-Neurofilament H & M (NF-H/NF-M), Phosphorylated antibody (clone SMI 310) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3: Acidic, 100X (Cat. No. 927601), the tissue was incubated with 0.5 µg/ml of the primary antibody for 60 minutes at room temperature. BioLegend's Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm
  • SMI-310_PURE_Neurofilament_Phosphorylated_Antibody_1_123118_updated.png
    IHC staining of purified anti-Neurofilament H & M (NF-H/NF-M), Phosphorylated antibody (clone SMI 310) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3: Acidic, 100X (Cat. No. 927601), the tissue was incubated with 0.5 µg/ml of the primary antibody for 60 minutes at room temperature. BioLegend's Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm
  • SMI-310_PURE_Neurofilament_Phosphorylated_Antibody_2_123118.png
    Western blot of purified anti-Neurofilament H & M (NF-H/NF-M), Phosphorylated antibody (clone SMI 310). Lane 1: Molecular weight marker; Lane 2: 20 µg of human brain lysate; Lane 3: 20 µg of mouse brain lysate; Lane 4: 20 µg of rat brain lysate. The blot was incubated with 5 µg/mL of the primary antibody overnight at 4°C, followed by incubation with HRP labeled goat anti-mouse IgG (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system.
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837703 25 µg £77
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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 regulate 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-L) runs at 68-70 kD; the medium or middle (NF-M) runs at about 145-160 kD; 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 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 glia (negative for NF).

Product Details
Technical Data Sheet (pdf)

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.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

IHC-P - Quality tested
WB - Verified
IHC-F, ICC - Reported in the literature, not verified in house
Recommended Usage

For immunohistochemistry, a concentration range of 0.5 - 10 μg/ml is suggested. For Western blotting, the suggested use of this reagent is 0.5 - 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) include: Western blotting4, immunocytochemistry2, immunohistochemical staining of frozen tissue sections1,3, and spatial biology (IBEX)5,6.

 

Clone SMI 310 reacts with an extensively phosphorylated epitope of neurofilament H and, to a much lesser extent, neurofilament M in most mammalian species. Phosphatase treatment of samples abolishes reaction with SMI 310. A very extensive degree of hyperphosphorylation of neurofilaments seems to be necessary for its reactivity.

Application References

(PubMed link indicates BioLegend citation)
  1. Dubourg O, et al. 2011. Acta Myol. 30(2):103. (IHC-F) PubMed
  2. Mizui T, et al. 2009. J Neurochem. 109(2):611. (ICC)
  3. Denlger-Crish C, et al. 2014. Front Neursci. 8:290. (IHC-F) PubMed
  4. Mulot SF, et al. 1994. FEBS Lett. 349(3):359. (WB)
  5. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  6. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
RRID
AB_2566638 (BioLegend Cat. No. 837703)
AB_2566638 (BioLegend Cat. No. 837704)

Antigen Details

Structure
The medium or middle neurofilament (NF-M) runs at 145-160 kD, and the heavy or highest neurofilament (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 while other cells and tissues are unreactive, except for peripheral axons.
Cell Type
Mature Neurons
Biology Area
Cell Biology, Neuroscience, Neuroscience Cell Markers
Molecular Family
Intermediate Filaments, Phospho-Proteins
Antigen References
  1. Siedler D, et al. 2014. Front Cell Neurosci. 8:429.

 

Gene ID
4744 View all products for this Gene ID
UniProt
View information about Neurofilament HM NF-H NF-M Phospho on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 3    Revision Date: 12/31/2018

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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