Vimentin (VIM) Antikörper

Antigen: Vimentin (VIM)

Synonyme: FLJ36605, MGC102095, cb28, MGC139781, VIM, vim, Vim, vime

Klonalität: Monoklonal (Vim 3B4)

Wirt: Maus

Applikation: Immunhistochemie (Paraffinschnitte) (IHC (p))

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370703

Produktbeschreibung

Weitere Bezeichnung: Vimentin

Immunogen: Vimentin isolated from bovine eye lens (3).

Isotyp: IgG2a, kappa chain  (Passende Sekundärantikörper)

Klon: Vim 3B4

Beschreibung: Vimentin is a 55 kDa intermediate filament (IF) protein, which form part of the cytoskeleton of vertebrate cells. Among the five classes* of IFs vimentin belongs to class III and is expressed in the cytoplasma of cells of mesenchymal origin (2). Being the predominant IF protein in mesenchymal cells vimentin was initially believed to be a useful marker for distinguishing non-epithelial from epithelial derived nepolasms. The co-expression of intermediate filaments, particularly vimentin and cytokeratin, has now been demonstrated in a variety of normal cells/tissues and in neoplastic lesions, necessitating the use of a panel of antibodies in differential tumour diagnosis (1). *Recently, an additional class (i.e. class VI) was created for nestin (2).

Spezifität: SDS-PAGE analysis of immunoprecipitates formed between the antibody and 35 S-labelled proteins from metabolically labelled human osteogenic sarcoma 4-998 cells shows reaction with a ~60 kDa polypeptide, corresponding to vimentin. In addition, a few bands of lower molecular weight were observed and may represent degradation products of vimentin (4). In Western blotting of cytoskeletal proteins from Xenopus laevis kidney epithelial cells and erythrocytes, the antibody labels a band of ~57 kDa corresponding to vimentin. No labelling was observed of other intermediate filaments (3). The antibody reacts with an epitope that has been localized to the coil 2 part of the vimentin rod domain. This epitope has been shown to encompass a region around residue 353 of the rod domain of mouse vimentin (5). As demonstrated by immunocytochemistry on frozen as well as formalin-fixed tissues, the antibody cross-reacts with the vimentin-equivalent protein in man, chicken, cow, dog, hamster, horse, monkey, rabbit and Xenopus laevis (3, 5-8).

Anwendungen

Applikationshinweise: Paraffin sections: The antibody can be used for labelling paraffin-embedded tissue sections fixed in formalin. Pre-treatment of tissues with proteinase K or heat-induced epitope retrieval is required. For heat-induced epitope retrieval, optimal results are obtained with DakoCytomation Target Retrieval Solution, high pH, code No. S3308, DakoCytomation Target Retrieval Solution, code No. S1700, 10 mmol/L citrate buffer, pH 6.0, or 10 mmol/L Tris buffer, 1 mmol/L EDTA, pH 9.0. The tissue sections should not dry out during the treatment or during the following immunocytochemical staining procedure. Frozen sections and cell preparations: The antibody can be used for labelling acetone fixed, frozen sections (3, 6, 7). Dilution: Monoclonal Mouse Anti-Vimentin, code No. M7020, may be used at a dilution range of 1:100-1:200 when applied on formalin-fixed, paraffin-embedded sections of malignant melanoma and using 20 minutes heat-induced epitope retrieval in DakoCytomation Target Retrieval Solution, High pH, code No. S3308, and 30 minutes incubation at room temperature with the primary antibody. Optimal conditions may vary depending on specimen and preparation method, and should be determined by each individual laboratory. The recommended negative control is DakoCytomation Mouse IgG2a, code No. X0943, diluted to the same mouse IgG concentration as the primary antibody. Unless the stability of the diluted antibody and negative control has been established in the actual staining procedure, it is recommended to dilute these reagents immediately before use, or dilute in DakoCytomation Antibody Diluent, code No. S0809. Positive and negative controls should be run simultaneously with patient specimen. Visualization: LSAB/HRP kit, code No. K0679, and EnVision/HRP kits, code Nos. K4004 and K4006, are recommended. For frozen sections and cell preparations, the DakoCytomation APAAP kit, code No. K0670, is a good alternative if endogenous peroxidase staining is a concern. Follow the procedure enclosed with the selected visualization kit. Automation: The antibody is well-suited for immunocytochemical staining using automated platforms, such as the DakoCytomation Autostainer. Performance characteristics: Cells labelled by the antibody display staining confined to the cytoplasm. Normal tissues: The antibody strongly labels fibrocytes, lipocytes, smooth muscle cells, vascular endothelial cells, periferal nerve (Schwann) cells, macrophages (including Kupffer cells) and myoepithelial cells of sweat and salivary glands.

Buffer: Monoclonal mouse antibody provided in liquid form purified from cell culture supernatant. In 0.05 mol/L Tris/HCl, 1% bovine serum albumin (BSA) and 15 mmol/L NaN 3, , pH 7.2.

Lagerung: Store at 2-8 °C. Precautions: 1. For professional users. 2. This product contains sodium azide (NaN 3 ), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used. 4. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin. 5. Unused solution should be disposed of according to local, State and Federal regulations.

Forschungsgebiet: Zell-/Gewebemarker, Zytoskelett

Beschränkungen: Für Forschungszwecke. Als CE-zertifizierter Antikörper in der Europäischen Union für In Vitro Diagnostik (IVD) zugelassen.

Publikationen

Publikationen: Borthwick, McIlroy, Gorowiec et al.: "Inflammation and epithelial to mesenchymal transition in lung transplant recipients: role in dysregulated epithelial wound repair." in: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, Vol. 10, Issue 3, pp. 498-509, 2010 (PubMed).

Klebe, Brownlee, Mahar et al.: "Sarcomatoid mesothelioma: a clinical-pathologic correlation of 326 cases." in: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, Vol. 23, Issue 3, pp. 470-9, 2010 (PubMed).

Hudelist, Huber, Auer et al.: "Administration of betaHCG leads to dose-dependent changes of gene expression signature of endometriotic stromal cells." in: Reproductive biomedicine online, Vol. 20, Issue 5, pp. 699-706, 2010 (PubMed).