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Alexa Fluor® 647 anti 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
Ave. Rating
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Product Citations
publications
SMI-35_A647_NeurofilamentH&M_Antibody_IHC_1_050818
IHC staining of Alexa Fluor® 647 anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated antibody (clone SMI 35) on formalin-fixed paraffin-embedded human brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3 (Cat. No. 927601), the tissue was incubated with 10 µg/ml of the primary antibody overnight at 4°C. The image was captured with a 40X objective. Scale bar: 50 µm
  • SMI-35_A647_NeurofilamentH&M_Antibody_IHC_1_050818
    IHC staining of Alexa Fluor® 647 anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated antibody (clone SMI 35) on formalin-fixed paraffin-embedded human brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3 (Cat. No. 927601), the tissue was incubated with 10 µg/ml of the primary antibody overnight at 4°C. The image was captured with a 40X objective. Scale bar: 50 µm
  • SMI-35_A647_NeurofilamentH&M_Antibody_IHC_2_050818
    IHC staining of Alexa Fluor® 647 anti-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 (Cat. No. 927601), the tissue was incubated with 5 µg/ml of the primary antibody overnight at 4°C. The image was captured with a 40X objective. Scale bar: 50 µm
  • SMI-35_A647_NeurofilamentH&M_Antibody_IHC_3_050818
    IHC staining of Alexa Fluor® 647 anti-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 (Cat. No. 927601), the tissue was incubated with 10 µg/ml of the primary antibody overnight at 4°C. The image was captured with a 40X objective. Scale bar: 50 µm
  • SMI-35_A647_NeurofilamentH&M_Antibody_IHC_4_050818
    IHC staining of Alexa Fluor® 647 anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated antibody (clone SMI 35) on formalin-fixed paraffin-embedded human brain tissue. Following antigen retrieval using Retrieve-All Antigen Unmasking System 3 (Cat. No. 927601), the tissue was incubated with 10 µg/ml of the primary antibody overnight at 4°C. The image was captured with a 10X objective. Scale bar: 100 µm
  • SMI-35_A647_NeurofilamentH&M_Antibody_IHC_5_050818
    IHC staining of Alexa Fluor° 647 anti-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 (Cat. No. 927601), the tissue was incubated with 10 µg/ml of the primary antibody overnight at 4°C. The image was captured with a 10X objective. Scale bar: 100 µm
  • SMI-35_A647_NeurofilamentH&M_Antibody_IHC_6_050818
    IHC staining of Alexa Fluor® 647 anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated antibody (clone SMI 35) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval Retrieve-All Antigen Unmasking System 3 (Cat. No. 927601), the tissue was incubated with 10 µg/ml of the primary antibody overnight at 4°C. The image was captured with a 10X objective. Scale bar: 100 µm
Compare all formats See Alexa Fluor® 647 spectral data
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835611 25 µg 104€
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835612 100 µg 259€
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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).

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 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 - 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 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
  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_2734602 (BioLegend Cat. No. 835611)
AB_2734602 (BioLegend Cat. No. 835612)

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: 1    Revision Date: 05.10.2018

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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