| Name: | Chicken Polyclonal to Vimentin |
| Immunogen: | Full length recombinant human vimentin protein , PROT-r-Vim, expressed in and purified from E. coli. expressed in and purified from E. coli. |
| HGNC Name: | VIM |
| UniProt: | P08670 |
| Molecular Weight: | 50kDa |
| Host: | Chicken |
| Isotype: | |
| Species Cross-Reactivity: | Human, rat, mouse, cow, pig, horse, chicken |
| RRID: | AB_2216401 |
| Format: | Concentrated IgY preparation in PBS plus 0.02% NaN3 |
| Applications: | WB, IF/ICC |
| Recommended Dilutions: | WB: 1:5,000. IF/ICC: 1:10,000. IHC not recommended |
| Storage: | Stable at 4°C for at least one year |
Immunofluorescent analysis of HeLa cell culture stained with chicken pAb to vimentin, CPCA-Vim, dilution 1:10,000 in green, and costained with EnCor mouse mAb to actin, MCA-5J11, dilution 1:500 in red. The blue is DAPI staining of nuclear DNA. The vimentin antibody stains the intermediate filament network while the actin antibody labels the submembranous cytoskeleton, stress fibers, and bundles of actin associated with cell adhesion sites.
Western blot analysis of tissue and cell lysates using chicken pAb to Vimentin, CPCA-Vim, dilution 1:5,000 in red. [1] protein standard (red), [2] rat whole brain lysate, [3] HeLa, [4] SH-SY5Y, [5] HEK293, and [6] NIH-3T3 cell lysates. CPCA-Vim binds to the vimentin protein showing a single band at ~50 kDa. The blot was simultaneously probed with mouse mAb to MAP2C/D, MCA-2C4, dilution 1:5,000 in green, revealing multiple bands around 280kDa that correspond to full length MAP2A/2B isotypes while the ~70kDa bands are MAP2C/D isotypes. MAP2 isotypes are seen only in extracts containing neuronal lineage cells.
Chicken Polyclonal Antibody to Vimentin
Cat# CPCA-Vim
$120.00 – $800.00
Vimentin is a protein subunit of the intermediate or 10nm filaments found in the cytoplasm of many cell types (1). Intermediate filaments are relatively stable fibrous components of cells which appear to have primarily a mechanical function. Many cell lines such as HEK293, HeLa, 3T3 and Cos cells contain prominent vimentin networks (1). Vimentin is a major protein of eye lens and cornea, and found generally in mesenchymal tissues in adult mammals. In the CNS it is found in endothelia and developing neurons, developing and some mature astrocytes, microglia, mature Bergmann glia and ependyma (2,3). Mutations in the vimentin gene may cause cataracts (4,5), and elevated levels of vimentin in blood samples are associated with onset of cancer (6,7). Vimentin levels increase in a variety of cell types as they become cancerous, suggesting that increase in expression of this protein is a useful diagnostic marker of the epithelial-mesenchymal transition, an important step in the metastasis of carcinoma cells (8).
The CPCA-Vim antibody can be used to study stem cells and generally to reveal the intermediate filament cytoskeleton. The antibody works well for IF and ICC, but is not recommended for IHC. The immunogen used to generate this antibody was full length recombinant human vimentin, PROT-r-Vim, expressed in and purified from E. coli. The antibody works well on all mammals tested to date, and it was generated in chicken by standard procedures and immunoglobulin was extracted from egg yolk. The resulting polyclonal antibody belongs to the IgY subclass, the chicken homolog of mammalian IgG and can be used in the same way, with the caveat that this type of antibody does not bind either Protein A or Protein G. Suitable second antibody reagents can be obtained from many vendors including ThermoFisher and Sigma-Aldrich. The same vimentin immunogen was used to produce two high quality epitope mapped monoclonal antibodies to vimentin MCA-2A52 and MCA-2D1, and also a rabbit polyclonal antibody RPCA-VIM. Mouse select image at left for larger view.
Neuromics hN2 are human neuronal lineage cells which originate from ArunA Biomedical, and were derived from the human embryonic cell line WA09. We obtained the hN2 Human Neurons Discovery kit from Neuromics, which arrived frozen and on dry ice. We fixed and stained them with our chicken polyclonal antibody to vimentin CPCA-Vim, in red. Islands of hN2 cells form after 4 days in culture forming beautiful flower like structures. Vimentin is a well established marker of early differentiating neuronal lineage cells, though it is found in many other kinds of cells. Merged from several images taken with a 10X objective lens, this image is also available as a high resolution poster. Blue staining is nuclear DNA.
1. Franke WW, et al. Different intermediate-sized filaments distinguished by immunofluorescence microscopy. PNAS 75:5034–8 (1978).
2. Dahl D, et al. Vimentin, the 57 000 molecular weight protein of fibroblast filaments, is the major cytoskeletal component in immature glia. Eur. J. Cell Biol. 24:191-6 (1981).
3. Shaw, G. et al. An immunofluorescence microscopical study of the neurofilament triplet proteins, vimentin and glial fibrillary acidic protein within the adult rat brain. Eur. J. Cell Biol. 26:68-72 (1981).
4. Muller M, et al. Dominant cataract formation in association with a vimentin assembly disrupting mutation. Hum. Molec. Genet. 18:1052-7 (2009).
5. Zhai Y, et al. Targeted exome sequencing of congenital cataracts related genes: broadening the mutation spectrum and genotype-phenotype correlations in 27 Chinese Han families. Sci. Rep. 7:1219 (2017).
6. Satelli A, Li S. Vimentin in cancer and its potential as a molecular target for cancer therapy. Cell Mol. Life Sci. 68:3033-46 (2011).
7. Wong KF, Luk JM. Discovery of lamin B1 and vimentin as circulating biomarkers for early hepatocellular carcinoma. Meth. Mol. Biol. 2909:295-310 (2012).
8. Jia X, et al. Vimentin-a potential biomarker for therapeutic efficiency of HAART. Acta Biochim. Biophys. Sin. (Shanghai) 6:1001-6 (2014).
