Preparation method of polyase marker antibody
A technology for labeling antibodies and polymerases, applied in the field of enzyme immunity, can solve the problems of many steps and complexity in the preparation process, and achieve the effects of simplified preparation process, high speed and high yield
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[0023] A method for preparing a polymerase-labeled antibody, comprising the steps of:
[0024] 1) Oxidation reaction: take 8mg horseradish peroxidase (HRP), add 8mmol NaIO 4 , reacted for 20 minutes; NaIO 4 The addition of (oxidizing agent) plays an oxidation role to obtain HRP containing aldehyde groups;
[0025] 2) polymerization to form polymerase (Poly-HRP): step 1) adding 2ml of mass concentration to the aldehyde-containing HRP obtained is the non-ionic surfactant TritonX-100 of 3%, and then 1M Na 2 CO 3 -NaHCO 3 Adjust the pH of the solution to 8.8-9.2 (optimally 9.0), then rotate and react overnight at 4°C, and react at room temperature for 1 hour the next day to obtain Poly-HRP; add non-ionic surfactant to form micellized coating;
[0026] 3) Purification: the polymerase obtained in step 2) was dialyzed in 10 mM phosphate physiological buffer solution (PBS) for 3 times to obtain a dialyzate;
[0027] 4) IgG connection: take out the dialyzate obtained in step 3) (t...
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