Immunostaining of a coronal section of the spinal cord of a rat given a midline C4 contusion injury three days previously. Sections were stained with <a href="http://encorbio.com/product/rpca-nf-l-ct" target="_blank" rel="noopener">RPCA-NF-L-ct</a> (red) and CPCA-NF-L-Degen in green. CPCA-NF-L-Degen stains prominent aggregates of material concentrated in the lateral funiculi and the dorsal columns but seen in lesser amounts throughout the section. These are degenerating and degenerated axons dam
Immunostaining of a coronal section of the spinal cord of a rat given a midline C4 contusion injury three days previously. Sections were stained with <a href="http://encorbio.com/product/rpca-nf-l-ct" target="_blank" rel="noopener">RPCA-NF-L-ct</a> (red) and CPCA-NF-L-Degen in green. CPCA-NF-L-Degen stains prominent aggregates of material concentrated in the lateral funiculi and the dorsal columns but seen in lesser amounts throughout the section. These are degenerating and degenerated axons dam

EnCor Biotechnology

Chicken Polyclonal Antibody to NF-L (Nfl, NEFL) DegenoTag™ Peptide Cat# CPCA-NF-L-Degen

$300.00
Description

      The CPCA-NF-L-Degen was raised against a proprietary recombinant immunogen containing amino acids 311-375 of the human NF-L sequence and is one of our :DegenoTag™" reagents which specifically binds to degenerated forms of NF-L. Full details of these findings are described in a peer-reviewed publication in Brain Communications. The antibody works well on western blots of a variety of species but binds only degenerated processes in sectioned material. We recommend using the antibody at 1:10,000 dilution for the specific visualization of degenerating and degenerated processes by IF/ICC. It also works well on paraffin embedded histological sections of rodent CNS tissues, including transgenic mouse models. This and other DegenoTag™ reagents can be used to identify degenerating and degenerated processes and also to monitor NF-L degradation in a variety of contexts. EnCor generated another polyclonal DegenoTag™ reagent made in rabbit RPCA-NF-L-Degen. We also generated several monoclonal antibodies specific for degenerated forms of NF-L, MCA-6H63, MCA-1D44 and MCA-1B11.

Amount: 100µL of 1mg/mL
Amount: 100µL of 1mg/mL
Immunostaining of a coronal section of the spinal cord of a rat given a midline C4 contusion injury three days previously. Sections were stained with RPCA-NF-L-ct (red) and CPCA-NF-L-Degen in green. CPCA-NF-L-Degen stains prominent aggregates of material concentrated in the lateral funiculi and the dorsal columns but seen in lesser amounts throughout the section. These are degenerating and degenerated axons damaged by the C4 lesion. The RPCA-NF-L-ct antibody binds the C-terminal "tail" region of NF-L which is absent or destroyed during degeneration, so the CPCA-NF-L-Degen positive profiles are largely negative for RPCA-NF-L-ct.
Western blot analysis of different tissue lysates using chicken pAb degenerated forms of NF-L, CPCA-NF-L-Degen dilution 1:10,000 in green: [1] protein standard, [2] rat brain, [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord, [6] cow spinal cord and [7] pig spinal cord. The strong band at about 68kDa corresponds to full length denatured NF-L protein. Lower molecular weight bands detected mainly in the spinal cord samples are proteolytic forms of NF-L.

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Name: CPCA-NF-L-Degen
Immunogen: Degenotag™ NF-L peptide
HGNC Name: NEFL
UniProt: P07196
Molecular Weight: 68-70kDa
Host: Chicken
Species Cross-Reactivity: Human, rat, mouse, pig, cow
RRID: AB_2923500
Format: Purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM azide
Applications: WB, IF/ICC, IHC
Recommended Dilutions: WB: 1:10,000-1:20,000. IF/ICC: 1:5,000-1:10,000. IHC: 1:5,000.
Storage: Stable at 4°C for one year, for longer term store at -20°C

       Neurofilaments are major components of neurons and their axons (1-5). We have recently developed a series of novel reagents which we call DegenoTag™ products. These are antibodies which recognize epitopes in a small segment of the neurofilament NF-L subunit which are normally not accessible to antibodies but which became available on degeneration. We propose that these epitopes are made accessible as a result of degeneration induced proteolysis, and in agreement with this hypothesis we could make previously negative control tissues become strongly DegenoTag™ antibody positive by treatment with proteases. In addition fresh CNS tissues did not stain with DegenoTag™ reagents except for a tiny minority of apparently spontaneously degenerating processes. In stark contrast tissues left to sit at room temperature for 4 hours were strongly reactive with DegenoTag™ reagents. We also discovered that our antibodies to the C-terminal of NF-L, such as our rabbit polyclonal RPCA-NF-L-ct and mouse monoclonal MCA-DA2. Our reagents can therefore be used to positively identify both healthy and degenerated processes



Chromogenic immunohistochemistry on a formalin fixed paraffin embedded spinal cord section of an ALS mouse model from the lab of Dr. David Borchelt (6). These heterozygous G85R-SOD1-YFP mice do not develop ALS unless they are primed by injection of mutant SOD1 aggregates, so this is a model of prion induction of pathogenesis. This mouse was injected with mutant SOD1 in the hind limb and degenerated processes were seen in the sciatic nerve, in fibers in the spinal cord and as shown here in the brain stem. Staining was performed with chicken pAb to degenerated neurofilament NF-L, CPCA-NF-L-Degen, dilution 1:10,000, detected with DAB (brown) using the ABC method and reagents with no antigen retrieval.Hematoxylin (blue) was used as the counterstain. Hematoxylin (blue) was used as the counterstain. Mouse select image for larger view.

1. Hoffman et al. Neurofilament gene expression:a major determinant of axonal caliber. PNAS 84:3472-6 (1987).
2. Perrot R, et al. Review of the Multiple Aspects of Neurofilament Functions, and their Possible Contribution to Neurodegeneration. Mol. Neurobiol. 38:27-65 (2008).
3. Lépinoux-Chambaud C. Eyer J. Review on intermediate filaments of the nervous system and their pathological alterations. Histochem. Cell Biol. 140:13-22 (2013).
4. Liu Q. et al. Neurofilamentopathy in Neurodegenerative Diseases. Open Neurol. J. 5:58–62 (2011).
5. Bacioglu M, et al. Neurofilament light chain in blood and CSF as marker of disease progression in mouse models and in neurodegenerative diseases. Neuron 91:56-66 (2016).
6. Shaw G, et al. Uman Type NF-L Antibodies Are Effective Reagents for the Imaging of Neurodegeneration. BioRχiv DOI 10.1101/2022.08.27.504533 (2022).
7. Norgren N, et al. Monoclonal antibodies selective for low molecular weight neurofilaments. Hybrid Hybridomics 21:53-59 (2002).

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