Phospho-NF-H Antibody NE14 / Phospho-Neurofilament Heavy Chain Antibody [clone NE14] (V2759)

Description

Phospho-NF-H Antibody NE14 / Phospho-Neurofilament Heavy Chain Antibody detects a phosphorylated form of neurofilament heavy chain (NF-H), a neuronal intermediate filament protein encoded by the NEFH gene. NF-H is one of the major neurofilament subunits and contributes to the specialized cytoskeletal architecture of neurons and axons. The protein contains an extended C-terminal domain rich in repeated lysine-serine-proline (KSP) motifs that undergo extensive phosphorylation in neuronal processes.

Neurofilaments are composed primarily of light, medium and heavy subunits, commonly designated NF-L, NF-M and NF-H. Together, these proteins form intermediate filament networks that provide structural support to axons and contribute to the regulation of axonal caliber. NF-H has an apparent molecular weight of approximately 200 kDa and is distinguished by its long C-terminal tail. Extensive phosphorylation of this region influences neurofilament organization, interactions between neighboring filaments and properties of the axonal cytoskeleton.

Clone NE14 recognizes a phosphorylation-dependent epitope within the KSP/KEP-rich C-terminal region of NF-H. Antibody binding is lost following alkaline phosphatase-mediated dephosphorylation of neurofilaments, supporting the phospho-specific recognition of this monoclonal antibody. This specificity makes NE14 useful for distinguishing phosphorylated NF-H in studies of neurofilament organization and neuronal differentiation.

Phosphorylated NF-H is particularly associated with mature axonal neurofilaments and is widely investigated in neuroscience, neuropathology and studies of neuronal injury. Changes in NF-H phosphorylation can accompany alterations in neurofilament transport, axonal organization and neuronal integrity. Phosphorylated neurofilament heavy chain is also studied in neurodegenerative and neuroinflammatory disorders and as a marker of axonal damage, providing a link between neurofilament post-translational modification and nervous system pathology.

Clone NE14 is a mouse monoclonal antibody developed against a neurofilament preparation from porcine spinal cord and has been widely used in neurofilament research. The antibody has been evaluated by Western blot, immunohistochemistry and flow cytometry and recognizes human, mouse and rat NF-H. NSJ Bioreagents supplies antibodies for neuroscience and phosphorylation research. A Phospho-NF-H Antibody can be used to investigate phosphorylated neurofilament heavy chain, axonal cytoskeleton organization and changes in neurofilament phosphorylation associated with neuronal biology and disease.

For additional research on NF-H expression, axonal structure and neurofilament biology, see our Neurofilament Heavy Chain Antibody / Axonal Cytoskeleton Marker page.

Application Notes

Optimal dilution of the Phospho-NF-H Antibody NE14 / Phospho-Neurofilament Heavy Chain Antibody should be determined by the researcher.

1. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM Citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 min
2. The prediluted format is supplied in a dropper bottle and is optimized for use in IHC. After epitope retrieval step (if required), drip mAb solution onto the tissue section and incubate at RT for 30 min.

Immunogen

Crude neurofilament preparation from porcine spinal cord was used as the immunogen for the Phospho-NF-H Antibody NE14.

Storage

Store the Phospho-NF-H Antibody NE14 at 2-8oC (with azide) or aliquot and store at -20oC or colder (without azide).

Alternate Names

Phospho-NF-H antibody, Phospho-Neurofilament Heavy Chain antibody, Phospho-NEFH antibody, Phosphorylated NF-H antibody, Neurofilament H Phospho antibody