Neurofilament NF-L, chicken polyclonal, Cat# CPCA-NF-L

Neurofilament NF-L, chicken polyclonal, Cat# CPCA-NF-L

View of mixed neuron/glial cultures stained with CPCA-NF-L (red) and EnCors rabbit antibody to phosphorylated NF-H (RPCA-NF-H, green). The NF-L protein is assembled into neurofilaments which are found throughout the axons, dendrites and perikarya of these cells. In contrast the phosphorylated NF-H has a more restricted expression pattern, being found only in developed axonal neurofilaments. Axonal profiles therefore are yellow since the red and green signals are superimposed. In contrast neurofilaments containing only NF-L appear red.

Western blots of whole protein extracts of rat spinal cord (lane 1), brain stem (lane 2), cerebellum (lane 3) and cerebral cortex (lane 4). Neurofilaments are concentrated in large projection axons and therefore NF-L is a more prominent component of spinal cord than cortical regions (see reference 1).

Molecular weight68 kDa by SDS-PAGE

Product name Anti-NF-L
Description Chicken polyclonal to Neurofilament Light chain, NF-L
Reference Code CPCA_NF-L
HGNC name NEFL
RRID# AB_2149931
Immunogen  Recombinant human NF-L protein
Isotype IgY
Concentration Antibody is supplied as an aliquot of concentrated IgY prep.
Species Reactivity Human, rat, mouse
Applications Western blot, ICC/IF, IHC,
Suggestions for use Western blot: 1:5,000. IF/ICC and IHC: 1:2,000.
Storage instructions Shipped on ice. Store at 4°C. For long term storage, leave frozen at -20°C. Avoid freeze / thaw cycles.

Neurofilaments can be defined as the intermediate or 10nm filaments found in specifically in neuronal cells. In the electron microscope neurofilaments appear as 10nm diameter fibres of indeterminate length which generally have fine wispy protrusions from their sides. They are found to be particularly abundant in axons of large projection neurons. They probably function to provide structural support for neurons and their synapses and to support the large axon diameters required for rapid conduction of impulses down axons. They are composed as a mixture of subunits which usually include the three neurofilament triplet proteins, known as NF-L, NF-M and NF-H. Neurofilaments may also include smaller amounts of peripherin, α-internexin, nestin and in some cases vimentin.

Antibodies to the various neurofilament subunits are very useful cell type markers since the proteins are among the most abundant of the nervous system, are expressed only in neurons, and are biochemically very stable. To raise this antibody, bovine intermediate filaments were prepared from spinal cords by the method of Delacourte et al.(2). Individual neurofilament subunits were purified by ion exchange chromatography on DEAE cellulose in 6M urea followed by preparative SDS-PAGE. To ensure greater specificity for NF-L, animals were boosted with recombinant mouse NF-L purified from bacteria. The HGNC name for this protein is NEFH.

This antibody was generated in chicken by standard procedures and immunoglobulin was extracted from egg yolk. The resulting polyclonal antibody belongs to the IgY subclass. This is the chicken homologue of mammalian IgG and can be used in the same general way, with the caveat that this type of antibody does not bind either Protein A or Protein G. The IgY preparation was made by chloroform delipidation of egg yolk followed by polyethylene glycol precipitation. The IgY total concentration is is 21.5 mg/ml in phosphate buffered saline plus 10mM sodium azide preservative.

References:

1. Shaw G, Yang C, Ellis R, Anderson K, Parker Mickle J, Scheff S, Pike B, Anderson DK and Howland DR. Hyperphosphorylated neurofilament NF-H is a serum biomarker of axonal injury. Biochem Biophys Res Commun. 336:1268-1277 (2005).

2.  Delacourte A, Filliatreau G, Boutteau F, Biserte G and Schrevel J. Study of the 10-nm-filament fraction isolated during the standard microtubule preparation. Biochem J. 191:543-6. (1980).

Download Adobe Acrobat format product data sheet: <http://encorbio.com/wp-content/uploads/2016/05/in-preparation.pdf> .

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