Name: | Chicken polyclonal antibody to phosphorylated NF-H |
Immunogen: | Native NF-H purified from bovine spinal cord. |
HGNC Name: | NEFH |
UniProt: | P12036 |
Molecular Weight: | 200-220kDa by SDS-PAGE |
Host: | Chicken |
Isotype: | |
Species Cross-Reactivity: | Human, rat, mouse, cow, pig, dog, horse |
RRID: | AB_2149761 |
Format: | Concentrated IgY preparation in PBS plus 0.02% NaN3 |
Applications: | WB, IF/ICC, IHC, ELISA |
Recommended Dilutions: | WB: 1:20,000-1:50,000. IF/ICC, IHC: 1:20,000. |
Storage: | Store at 4°C. |
Chicken Polyclonal Antibody to NF-H
Cat# CPCA-NF-H
$150.00 – $1,000.00
Neurofilaments are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H, though other proteins may also be present. NF-H is the neurofilament high or heavy molecular weight polypeptide and runs on SDS-PAGE gels at 200-220 kDa, with some variability across species boundaries. The protein is in reality much smaller in molecular size, about 110kDa (1,2). The unusual SDS-PAGE mobility is due partly to a very high content of charged amino acids, particularly glutamic acid rich regions, and the non-phosphorylated form runs on SDS-PAGE at about 160kDa. The predominant type of NF-H is the axonal form which is heavily serine phosphorylated on 40 or more tandemly repeated lysine-serine-proline (KSP) containing peptides (3-5). The phosphorylation of these peptides results in considerable further retardation on SDS-PAGE gels, so the heavily phosphorylated axonal form runs at 200-220kDa with some species variability. Antibodies to NF-H are useful for identifying axonal processes in tissue sections and in culture. NF-H antibodies can also be useful in visualizing neurofilament accumulations seen in many neurological disorders, such as Amyotrophic Lateral Sclerosis (also known as Lou Gehrig’s disease), Alzheimer’s disease and following traumatic injury. The phosphorylated axonal form of NF-H, usually referred to as pNF-H, can be detected in blood and CSF following a variety of damage and disease states resulting in axonal compromise, and antibodies such as this can be used to used to quantify such ongoing axonal loss (e.g. 6-8).
The CPCA-NF-H antibody was raised against biochemically isolated NF-H purified from bovine spinal cord (9). This preparation is dominated by axonal forms of NF-H which are heavily phosphorylated on the multiply repeated NF-H KSP type sequences, and this antibody reacts very strongly with these phosphorylated repeats. Reactivity with non-phosphorylated KSP sequences is orders of magnitude weaker, similar to other characterized antibodies to NF-H (5). In most species there is some cross-reactivity with the phosphorylated KSP sequences found in the related neurofilament subunit NF-M which are similar but not identical to those of NF-H. The antibody recognizes phosphorylated NF-H strongly in all mammals tested to date and also in chicken and has an unusually high titer of 1:20,000 or higher. The antibody is widely used and sold through many vendors, see for example the results of Google Scholar search for CPCA-NF-H. We also market mouse monoclonal antibodies MCA-NAP4, MCA-9B12 and MCA-AH1 and a rabbit polyclonal antibody RPCA-NF-H all of which have similar specificity to CPCA-NF-H. Mouse select image at left for larger view.
Chromogenic immunostaining of a formalin fixed paraffin embedded rat cerebellum section with chicken pAb to NF-H, CPCA-NF-H, dilution 1:10,000, detected in DAB (brown) following the ABC method. Hematoxylin (blue) was used as the counterstain. The NF-H antibody labels axonal profiles of basket cells surroundinf Purkinje neurons and processes from other types of neuron. Mouse select image for larger view.
This antibody has been widely used and marketed through most major antibody suppliers for many years. We have also sold it directly, see references in peer reviewed publications for the reagent obtained from EnCor here. A considerable literature now shows that detection of pNF-H in plasma, serum and CSF is a useful surrogate marker of axonal loss resulting from CNS damage disease (4).
The same antibody in the antigen affinity purified form has been used as a very effective ELISA capture reagent and is a component of our ELISA-pNF-H-v1 kit.
Peer reviewed publications using this antibody
1. Shields SD. et al. Nav1.8 expression is not restricted to nociceptors in mouse peripheral nervous system. Pain 153:2017-30 (2012).
Also check out this Google Scholar search; here.
1. Perrot R, et al. Review of the Multiple Aspects of Neurofilament Functions, and their Possible Contribution to Neurodegeneration. Mol. Neurobiol. 38:27-65 (2008).
2. Lépinoux-Chambaud C. Eyer J. Review on intermediate filaments of the nervous system and their pathological alterations. Histochem. Cell Biol. 140:13-22 (2013).
3. Sternberger LA, Sternberger NH. Monoclonal antibodies distinguish phosphorylated and nonphosphorylated forms of neurofilaments in situ. PNAS
80:6126-30 (1983).
4. Julien JP, Mushynski WE. Multiple phosphorylation sites in mammalian neurofilament polypeptides. J. Biol. Chem. 257:10467-70 (1982).
5. Lee VM, et al. Identification of the major multiphosphorylation site in mammalian neurofilaments. PNAS 85:1998-2002 (1988).
6. Shaw G, et al. Hyperphosphorylated neurofilament NF-H is a serum biomarker of axonal injury. Biochem. Biophys. Res. Commun. 336:1268-77 (2005).
7. Boylan et al, Immunoreactivity of the phosphorylated axonal neurofilament H subunit (pNF-H) in blood of ALS model rodents and ALS patients: evaluation of blood pNF-H as a potential ALS biomarker. J. Neurochem. 111:1182-91 (2009).
8. Shaw G. The Use and Potential of pNF-H as a General Blood Biomarker of Axonal Loss: An Immediate Application for CNS Injury. In: Kobeissy FH, editor. Brain Neurotrauma: Molecular, Neuropsychological, and Rehabilitation Aspects. CRC Press/Taylor & Francis; 2015. Chapter 21 .
9. Delacourte A, et al. Study of the 10-nm-filament fraction isolated during the standard microtubule preparation. Biochem. J. 191:543-6 (1980).
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Contact info
EnCor Biotechnology Inc.
4949 SW 41st Boulevard, Ste 40
Gainesville
Florida 32608 USA
Phone: (352) 372 7022
Fax: (352) 372 7066
E-mail: admin@encorbio.com