The binding region of this antibody has been characterized by Bohn et al. (1992). According to these authors, the epitope has been localized on the alphahelical part of vimentin (rod domain coil 2). Due to an aa substitution at position of aa353 in murine vimentin (that could explain for the weak crossreaction of the antibody with murine vimentin) they were able to narrow down the binding region around position 353. These findings were confirmed by truncation mutagenesis experiments using human vimentin (Rogers et al., 1995). Applications: Suitable for use in Western Blot, FLISA, Immunofluorescence Microscopy and Immunohistochemistry. Other applications not tested. Recommended Dilution: Immunohistochemistry (frozen and paraffin-embedded tissue and cytological material): 1:10 with PBS, pH 7.4, Incubation Time 1 hr at RT, extended with paraffin. Protease pretreatment is required prior to antibody application. Optimal dilution determined by the researcher. Positive Controls: Cultured RD cells, glioma cells, fibroblasts (SV-80), MDCK. Tumors specifically detected: sarcoma (including myosarcoma), lymphoma, melanoma. Storage and Stability: Store product at 4C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20C. Aliquots are stable at -20C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: FITC conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
