The digoxin system is over provided as another non-isotopic labeling method, which is detected by coupling an anti-digoxin antibody to one or more fluorescent dyes or enzymes, or by indirect immunofluorescence.

Operation method

Basic methods

Materials and Instruments

DNA
dTTP dUTP EDTA SDS
Water Bath Incubator

Move

1. Establish a standard 100 μl reaction system with 10 μl of 10× digoxigenin-11-dUTP/dTTP reservoir, with no change in the amount of DNAzyme Ⅰ enzyme, and incubate at 15°C for 2 h.

2. Take a small portion in microgel for electrophoresis to detect the probe size.

3. Continue the reaction until a DNA probe of appropriate size is obtained.4. Add 2 μl of 0.5 mol/l EDTA and 1 μl of 10% SDS and heat at 68°C for 10 min to terminate the reaction.

5. Separate the probe with a G-50 centrifugal column.6. Determination of digoxin-labeled probes by colorimetric or chemiluminescent methods.

7. Replacement of streptavidin-alkaline phosphatase cross-links with alkaline phosphatase-coupled anti-digoxin antibodies.

8. Detection of digoxigenin incorporation efficiency using digoxigenin-labeled labeled DNA or labeled digoxigenin probe as a control.

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