| 摘要 |
<p>A method for the preparation of tyramide conjugates is provided which can be utilized in the CARD amplification system. The method comprises reacting a suitable ester of a labeling compound with tyramine in a substantially water-free organic solvent at a constant pH of about 7 to about 9.5, preferably from 7.0 to 8.0. The reaction is suitably carried out in a buffered medium. Preferably, the NHS ester is applied of biotin, digoxigenin, dinitrobenzene, trinitrobenzene or the fluorochromes fluorescein, rhodamine, cyanine, coumarin and BODIPY dyes. The invention provides an easy and fast method to prepare a variety of known and new tyramide conjugates in bulk amounts at high purity and at relatively low cost. The tyramide conjugates thus obtained can be suitably applied in affinity assays including immunohistochemistry and in situ hybridization ISH procedures to detect with high efficiency protein epitopes or repetitive and single-copy DNA target sequences in cell preparations.</p> |
