Catalogue# MCA-DA2: Neurofilament NF-L Monoclonal Antibody Clone DA2
The Immunogen: 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. NF-L is the neurofilament light or low molecular weight polypeptide and runs on SDS-PAGE gels at about 68kDa. Antibodies to NF-L are useful for identifying neuronal cells and their processes in tissue sections and in tissue culture. NF-L antibody can also be useful in the diagnostics of neurofilament accumulations seen in many neurological diseases, such as Lou Gehrig's disease and Alzheimer's disease. Mutations in the protein coding region of the human NF-L gene cause some forms of Charcot-Marie-Tooth disease (1). This antibody has been sold by Chemicon and Millipore for many years under the catalogue number Mab1615, and has been used in nmerous publications (2-6). The same antibody is also marketed, more expensively, by Abcam, Novus and many other companies.
Left: Rat spinal cord homogenate showing the major intermediate filament proteins of the nervous system (lane 1). The remaining lanes show blots of this material stainted with various antibodies including MCA-DA2 (lane 4). Right: Cells grown from adult rat brain using method described by Evans et al. (2). Large cell in middle is stained with mouse monoclonal to NF-L clone DA2 (green). Another type of neuronal lineage cell was stained with rabbit polyclonal to alpha-internexin RPCa-a-Int (red). These cells were mitotic but had several characteristics of neurons.
Antibody characteristics: MCA-DA2 was raised against a preparation of enzymatically dephosphorylated pig neurofilaments including NF-L, NF-M and NF-H. Screening was by ELISA on the immungen followed by immunofluorescence microscopy. Clones which revealed strong staining were further characterized biochemically. MCA-DA2 is a IgG1 class antibody with a k light chain. It reacts with NF-L from human, cow, pig, mouse, rat and all other mammals, and also recognizes avian NF-L. It is strong and clean on western blots and works well on frozen sections, cells in tissue culture and on mildly formalin fixed histological sections.
Suggestions for use: For immunofluorescence, use MCA-DA2 diluted 1/500. For western blots try MCA-DA2 diluted 1/10,000 to 1:20,000. Store at 4°C short term or -20°C long term. Avoid repeated freezing and thawing.
Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.
References:
1. Mersiyanova IV, Perepelov AV, Polyakov AV, Sitnikov VF, Dadali EL, Oparin RB, Petrin AN and Evgrafov OV. A new variant of Charcot-Marie-Tooth disease type 2 is probably the result of a mutation in the neurofilament-light gene. Am. J. Hum. Genet. 67:37-46, 2000.
2. Edson KJ, Linck RW, Letourneau PC. Identification of a tektin-like protein associated with neurofilaments in the developing chick nervous system. J Neurosci Res. 30:105-15 1991.
3. Bongarzone ER, Foster L, Byravan S, Casaccia-Bonnefil P, Schonmann V, Campagnoni AT. Two neuronal cell lines expressing the myelin basic protein gene display differences in their in vitro survival and in their response to glia. J Neurosci Res. 54:309-19 1998.
4. Evans J. et al. Characterization of mitotic neurons derived from adult rat hypothalamus and brain stem. J. Neurophysiol. 87:1076-85 2002.
5. Shaw G, Morse S, Ararat M, Graham FL. Preferential transformation of human neuronal cells by human adenoviruses and the origin of HEK 293 cells. FASEB J. 16:869-71 2002.
6. Zhang L, Gavin T, DeCaprio AP, LoPachin RM. Gamma-diketone axonopathy: analyses of cytoskeletal motors and highways in CNS myelinated axons. Toxicol Sci. 117:180-9 2010.
7. Paulussen M, Jacobs S, Van der Gucht E, Hof PR, Arckens L. Cytoarchitecture of the mouse neocortex revealed by the low-molecular-weight neurofilament protein subunit. Brain Struct Funct. 216:183-99 2011.
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Availability and Price: Available for shipping now, $200 US per aliquot of 100 microliters of ascites fluid, enough for hundreds of experiments. For order form press here
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