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Monoclonal antibodies against isoprenoid compounds

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Wanke M., Porebska P. A., Porebska A., Walajtys-Rode W.-R. E., Walajtys-Rode E.
Biotechnologia
|
2001
|
issue 3
169-177
EN
Monoclonal antibodies against isoprenoid compounds were obtained by in vivo immunization of Balb/c mice with UQ-10 (ubiquinone-10) incorporated into cholesterol rich particles containing metylated bovine serum albumin (CRP-MBSA) in order to induce a strong immune response. The specificity of antibodies was evaluated by a modified dot-blot technique using Hybond-N+ membrane (Amersham). Selected antibodies III-5 (IgG3), IV-3 (IgG2b) and VII-11 (IgM) were capable of recognizing different isoprenoid compounds: ubiquinone, plastoquinone, solanesol, dolichol, -sistosterol but did not react with phospholipids, triacylglycerols or fatty acids. These antibodies may be used in immunoblotting for identifying isoprenoids (e.g. ubiquinone) in partially purified plant extracts after TLC chromatography on plastic RP-18 plates (Merck). Immunodetection of isoprenoids was performed using secondary goat anti-mice immunoglobin antibodies conjugated with biotin, extravidine-AP and BCIP/NBT as substrate
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