The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Reconstitution to filaments:
After vimentin is dissolved in 9.5M urea buffer, protofilaments and filament complexes are obtained by dialyzing the resulting polypeptide solution stepwise to a concentration of 4M urea and then to low salt condition (50mM NaCl, 2mM dithiothreitol, 10mM Tris-HCl, pH 7.4). For immunization purposes, the solution can be further dialyzed against PBS (phosphate buffered saline, e.g. Dulbecco's PBS).
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Preparation and Storage
Stability and Storage
Shipped at 4°C. Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Reconstitute in sterile 18MO-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Upon reconstitution ab73843 should be stored at 4°C between 2-7 days and for future use below -18°C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Please prevent freeze-thaw cycles.
Epididymis luminal protein 113
Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally. Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.
Highly expressed in fibroblasts, some expression in T- and B-lymphocytes, and little or no expression in Burkitt's lymphoma cell lines. Expressed in many hormone-independent mammary carcinoma cell lines.
Involvement in disease
Belongs to the intermediate filament family.
The central alpha-helical coiled-coil rod region mediates elementary homodimerization. The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.
Filament disassembly during mitosis is promoted by phosphorylation at Ser-55 as well as by nestin (By similarity). One of the most prominent phosphoproteins in various cells of mesenchymal origin. Phosphorylation is enhanced during cell division, at which time vimentin filaments are significantly reorganized. Phosphorylation by PKN1 inhibits the formation of filaments. Phosphorylated at Ser-56 by CDK5 during neutrophil secretion in the cytoplasm. Phosphorylated by STK33. O-glycosylated during cytokinesis at sites identical or close to phosphorylation sites, this interferes with the phosphorylation status. S-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex.
Western blot - Recombinant Human Vimentin protein (ab73843)
Ab45939 recognizes full length recombinant Human vimentin (ab73843) which has an expected molecular weight of 54 kDa.
This product has been referenced in:
Frescas D et al. Senescent cells expose and secrete an oxidized form of membrane-bound vimentin as revealed by a natural polyreactive antibody. Proc Natl Acad Sci U S A114:E1668-E1677 (2017).
ELISA, Sandwich ELISA
Read more (PubMed: 28193858) »
Twine NA et al. Transcription factor ZNF25 is associated with osteoblast differentiation of human skeletal stem cells. BMC Genomics17:872 (2016).
Read more (PubMed: 27814695) »