Alexa Fluor® 594 anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated Antibody

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
Isotype
Mouse IgG1, κ
a-SMI-35_A594_Neurofilament_2_IF_101917.png
Immunofluorescence staining of anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated (SMI 35) conjugated to Alexa Fluor® 594 on formalin-fixed, paraffin-embedded rat brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking Solution, the tissue was incubated with the Alexa Fluor® 594-conjugated antibody at 5µg/mL for one hour at room temperature. The image was captured with a 40X objective.
  • a-SMI-35_A594_Neurofilament_2_IF_101917.png
    Immunofluorescence staining of anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated (SMI 35) conjugated to Alexa Fluor® 594 on formalin-fixed, paraffin-embedded rat brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking Solution, the tissue was incubated with the Alexa Fluor® 594-conjugated antibody at 5µg/mL for one hour at room temperature. The image was captured with a 40X objective.
  • b-SMI-35_A594_Neurofilament_1_IF_101917.png
    SH-SY5Y neuroblastoma cells were fixed with 4% paraformaldehyde, and then permeabilized with 0.1% Triton x100 for 20 minutes. Cells were blocked with 2% Normal Goat Serum for 30 minutes at room temperature, then incubated with Alexa Fluor® 594 conjugated anti-NF-H/M Hypophosphorylated antibody at 5µg/mL for 3 hours at room temperature. Nuclei were stained with DAPI (bottom panel, blue). Images were captured with a 40X objective.
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835609 25 µg $129.00
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835610 100 µg $323.00
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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
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.
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 - Verified
Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration of 5.0 µg/mL is suggested. For immunocytochemistry, a concentration range of 5.0 - 10 μg/mL is recommended. 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 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
RRID
AB_2687299 (BioLegend Cat. No. 835609)
AB_2687299 (BioLegend Cat. No. 835610)

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: 3    Revision Date: 05/04/2022

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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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