Catalogue MCA-2D1: Mouse Monoclonal Antibody to Vimentin-VIM
The Immunogen: Vimentin is a protein subunit of 10nm or intermediate filaments, which are major components of the cytoskeleton in most cell types. Vimentin is the major intemediate filament subunit found in mesenchymal cells, and was first named and characterized in a collaborative study from the labs of German scientists Werner Franke and Klaus Weber (1). The name derives from the Latin "Vimentum", meaning arrays of flexible rods such as in lattices, filigrees and wicker-work, which describes the intermediate filament network quite well. Vimentin is also found in many cell types in tissue culture, most notably fibroblasts, and in developing neuronal and astrocytic precursor
cells in the central nervous system. Many cell lines such as Hek293, HeLa, 3T3 and Cos cells contain prominent vimentin networks. Vimentin frequently forms copolymers
with other intermediate filament proteins, such as GFAP (in astrocytes, ependymal cells and neural stem cells), with desmin (in muscle and endothelial cells) and neurofilament proteins
(in developing neurons). A E151K point mutation in the vimentin gene was shown to be causative of an autosomal dominant pulverulent cataract disease, but so far only in a single patient (2). Vimentin is a major protein of eye lens and cornea, and this mutation renders the molecule unable assemble into normal 10nm filaments. Antibodies to vimentin are useful in studies
of stem cells and generally to reveal the filamentous cytoskeleton. The
immunogen used to generate our antibody was recombinant human vimentin expressed in and purified
from E. coli. The same immunogen was used to produce our other monoclonal antibody to vimentin MCA-2A52. We also market a very popular chicken polyclonal antibody to vimentin, CPCA-Vim. The HGNC name for this protein is VIM.
Left: Western blot of crude extract of the human carcinoma HeLa cell line. Lane 18 was probed with MCA-2D1 antibody. Note the strong clean band at the expected molecular weight of 50kDa. Lane 17 was probed with another monoclonal antibody to vimentin, our MCA-2A52 clone. Lane 15 was probed with our antibody to actin, giving an SDS-PAGE molecular weight of 42kDa, and lane 16 with MCA-3G12, our antibody to 14-3-3η (14-3-3 eta), which has an SDS-PAGE molecular weight of 28kDa. Right: View of mixed neuron/glial cultures stained with MCA-2D1 (green) our our rabbit antibody to GFAP antibody (RPCA-GFAP, red). Vimentin is expressed alone in fibroblastic and endothelial cells, which are the flattened cells in the middle of the image which appear green. Astrocytes may express primarily GFAP, or GFAP and vimentin, and so appear red (GFAP only) or golden yellow (GFAP and Vimentin). In cells which express both GFAP and vimentin, the two protein assemble to produce heteropolymer filaments.
Antibody characteristics: MCA-2D1 is a mouse IgG2a class antibody with a k light chain. MCA-12D1 is known to react with vimentin from human, cow, pig, mouse, rat and other mammals. Since vimentin is highly conserved, it is likely that the antibody is effective on other species also.
Suggestions for use: The antibody solution is affinity purified from tissue culture supernatant and is at concentration of 1mg/ml in phosphate buffered saline. The antibody solution can be used at dilutions of at least 1:1,000 in immunofluorescence experiments. In western blotting using chemiluminescence it can be used at dilutions of 1:10,000 or lower. Antibody preparation contains 10mM sodium azide preservative (Link to http://www.encorbio.com/MSDS/azide.htm for Material Safety Data Sheet). Avoid repeated freezing and thawing, store at 4°C or -20°C.
UniProt Link: here
Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.
References:
1. Franke, W. W., Schmid, E., Osborn, M. and Weber, K. Different intermediate-sized filaments distinguished by immunofluorescence microscopy. Proc. Natl. Acad. Sci. USA 75:5034–5038 (1978).
2. Muller, M., Bhattacharya, S. S., Moore, T., Prescott, Q., Wedig, T., Herrmann, H., Magin, T. M. Dominant cataract formation in association with a vimentin assembly disrupting mutation. Hum. Molec. Genet. 18:1052-1057 (2009).
Availability and Price: Available for shipping now, $200 US per aliquot of 100 microliters of purified antibody at a concentration of 1 mg/ml, enough for hundreds of experiments. For order form press here
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