Rat mixed neuron/glial cultures stained with CPCA-Peri, (green channel) and the EnCor rabbit polyclonal antibody to the neurofilament subunit α-internexin RPCA-a-Int (red channel). These cultures contain mostly neurons which are rich in α-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 α-internexin, the green and red signals superimpose to produce a golden yellow cell. The blue signal is a DNA stain and reveals the nuclei of neurons and non-neuronal cells.
|These are Coomassie Brilliant Blue stained whole protein extracts of adult rat cortex (lane 1), brain stem (lane 2), cerebellum (lane 3), and spinal cord (lane 4) separated on 8% SDS-PAGE. The major neurofilament subunits are indicated by “NF-L”, “NF-M” and ”NF-H”, spectrin/fodrin by “S/F” and Microtubule associated protein 2 by “MAP2”. Right panel is a western blot of similar preparations in the reverse orientation, where lane 1 is spinal cord, lane 2 is brain stem, lane three is cerebellum and lane 4 is cortex. This blot if from a 6% SDS-PAGE and processed for immunoblotting with CPCA-Peri at a dilution of 1:10,000. A clear band running at 57kDa apparent size is seen in the spinal cord and brain stem lanes, but not in the other regions, in which peripherin is a very minor component. The position of this band on the Coomassie stained preparation is a little higher on the 8% gel and is indicated by “Peri” in the left panel.
||Chicken polyoclonal to Anti-Peripherin, PRPH
||Recombinant rat peripherin expressed in and purified from E. coli
||Antibody is supplied as an aliquot of concentrated IgY prep.
||Western blot, ICC/IF, IHC
|Suggestions for use
||Western blot: 1:20,000. ICC/IF and IHC: 1:2,000.
||Shipped on ice. Store at 4°C. For long term storage, leave frozen at -20°C. Avoid freeze / thaw cycles.
Peripherin is a ~57kDa intermediate filament subunit found initially in sensory neurons of the peripheral nervous systems, which gives the protein its name (1). 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,(PRPH2) 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.
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 26 mg/mL. Suitable second antibody reagents can be obtained from many vendors including Molecular Probes and Sigma-Aldrich.
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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