Anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated Antibody (Previously Covance catalog# SMI-35R)

Pricing & Availability
Clone
SMI 35 (See other available formats)
Regulatory Status
RUO
Other Names
Neurofilament heavy polypeptide, NF-H, 200 kD neurofilament protein, neurofilament triplet H protein
Previously
Covance Catalog# SMI-35R
Isotype
Mouse IgG1, κ
SMI-35_Purified_NF-H_Antibody_1_101618
IHC staining of Neurofilament H & M (NF-H/NF-M), Hypophosphorylated antibody (clone SMI 35) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3: Acidic, 100X (Cat. No. 927601), the tissue was incubated with a 1:2000 dilution of the primary antibody overnight at 4°C. 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 20X objective. Scale bar: 50 µm
  • SMI-35_Purified_NF-H_Antibody_1_101618
    IHC staining of Neurofilament H & M (NF-H/NF-M), Hypophosphorylated antibody (clone SMI 35) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3: Acidic, 100X (Cat. No. 927601), the tissue was incubated with a 1:2000 dilution of the primary antibody overnight at 4°C. 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 20X objective. Scale bar: 50 µm
  • SMI-35_Purified_NF-H_Antibody_2_101618
    IHC staining of Neurofilament H & M (NF-H/NF-M), Hypophosphorylated antibody (clone SMI 35) 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 a 1:2000 dilution of the primary antibody overnight at 4°C. 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-35_Purified_NF-H_Antibody_3_101618
    Western blot of Neurofilament H & M (NF-H/NF-M), Hypophosphorylated antibody (clone SMI 35). 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 a 1:5000 dilution 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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835601 100 µL $375.00
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835602 500 µL $673.00
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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).

Technical data sheet

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Preparation
Ascites
Concentration
The concentration is not quantified as this product is sold as undiluted crude mouse ascites fluid. The concentration might vary from lot-to-lot and an estimated concentration would be 1-3 mg/ml.
Storage & Handling
Store at -20°C. Upon initial thawing, apportion into working aliquots and store at -20°C. Avoid repeated freeze-thaw cycles to prevent denaturing the antibody. For long-term storage, keep the antibody at -80°C.
Application

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

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 1:1000 - 1:2000 µg/ml is suggested. For Western blotting, the suggested use of this reagent is 1:1000 - 1:5000 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, immunofluorescence staining, and spatial biology (IBEX)6,7.

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
  6. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  7. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Chung HL, et al. 2020. Neuron. 106(4):589-606. PubMed
  2. Das S, et al. 2020. Commun Biol. 0.354166667. PubMed
  3. Lu C, et al. 2012. PLoS One. 7:e40998. PubMed
  4. Neil S, et al. 2017. Brain Behav Immun. 10.1016/j.bbi.2017.02.018. PubMed
  5. Steinacker P, et al. 2011. PLoS One. 6:e23600. PubMed
RRID
AB_2565349 (BioLegend Cat. No. 835601)
AB_2565349 (BioLegend Cat. No. 835602)

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, 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 HM NF-H NF-M Phospho on UniProt.org
Go To Top Version: 4    Revision Date: 10/16/2018

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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.
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