The Immunogen:Peripherin is a ~57kDa intermediate filament subunit found initially in sensory neurons of the peripheral nervous systems, which gives the protein its name (1). The HGNC name for this protein is PRPH. Subsequently peripherin was found in some sensory and other neurons of the central nervous system and also in rat pheochromacytoma PC12 cells (2,3). Peripherin is also expressed in certain neuroendocrine tumors and in the insulin producing cells of the pancreas. Peripherin belongs to the Class III family of intermediate filament subunits which also includes vimentin, glial fibrillary acidic protein (GFAP) and desmin. Antibodies to peripherin can be used in identifying, classifying, and studying neurons throughout the nervous system. Peripherin is also a good diagnostic marker for ballooned axons seen in Lou Gehrig's disease (Amyotrophic lateral sclerosis) and some neuronally derived tumors. Autoantibodies to peripherin are frequently seen in the sera of patients with diabetes. Peripherin is not related to peripherin/RDS, a protein of the photoreceptor outer membrane mutations of which are causative of certain forms of slow retinal degeneration. The characterization of an antibody produced in the same way as CPCA-Peri, but in rabbit, has been published (4). The HGNC name for this protein is PRPH.

Figures: Blot in preparation. Right shows rat mixed neuron/glial cultures stained with CPCA-Peri, our chicken polyclonall antibody to peripherin (green channel) and EnCors rabbit antibody to the neurofilament subunit alpha-internexin RPCA-a-Int (red channel). These cultures contain mostly neurons which are rich in alpha-internexin, and a subgroup which have a large amount of peripherin also, such as the prominent cell in the middle of the micrograph. Since this cell expresses large amounts of both peripherin and alpha-internexin, the green and red signals superimpose to produce a golden yellow cell. Blue is a DNA stain.

Antibody Characteristics: 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 total concentration is is 26mg/ml. Suitable second antibody reagents can be obtained from many vendors including Molecular Probes and Sigma-Aldrich. Store at 4°C or -20°C. Avoid repeat freezing and thawing. Press here to download a material safety data sheet (MSDS) for sodium azide. Store at 4°C or -20°C. Avoid repeat freezing and thawing.

Suggestions for use: The IgY solution can be used at dilutions of at least 1:2,000 in immunofluorescence experiments. In western blotting using chemiluminescence it can be used at dilutions of 1:10,000 or lower. The solution has a total protein concentration of 21.5 mg/ml in phosphate buffered saline plus 10mM sodium azide.

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References:

1. Portier MM, de Néchaud B, Gros F. Peripherin, a new member of the intermediate filament protein family. Dev Neurosci. 6:335-44 (1984)

2. Troy CM, Brown K, Greene LA, Shelanski ML. Ontogeny of the neuronal intermediate filament protein, peripherin, in the mouse embryo. Neuroscience. 36:217-37 (1990).

3. Aletta JM, Angeletti R, Liem RK, Purcell C, Shelanski ML, Greene LA. Relationship between the nerve growth factor-regulated clone 73 gene product and the 58-kilodalton neuronal intermediate filament protein (peripherin).J Neurochem. 51:1317-20 (1988).

4. Errante LD, Wiche G and Shaw G. Distribution of plectin, an intermediate filament-associated protein, in the adult rat central nervous system J. Neurosci. Res. 37:515-528 (1994).

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