Fundamental
The dot gold immunofiltration assay is based on the speckle immunofiltration assay, which uses a colloidal gold label instead of an enzyme to eliminate the step of substrate color development. The test method is to use a nitrocellulose membrane (NC membrane) as a carrier, and to add a reagent and a sample to the membrane, and gradually react by diafiltration. The whole process can be completed in a few minutes, with positive results showing red spots on the membrane.
Reagents and materials
1. Percolation device: It is a plastic small box (4x3x0.6cm), divided into two layers, the bottom and the cover. The center of the cover has a small hole with a diameter of 0.5cm. The bottom of the box is filled with a pad with strong water absorption, and under the cover hole, a piece of NC film is placed next to the pad. Close the lid, which is the percolation device. Add 1-2 ml of specific antibody (primary antibody) or antigen to the NC membrane in the well, dry naturally at room temperature, and store in a sealed plastic bag with desiccant.
2. Blocking solution: Washing solution containing 0.2% BSA and 0.05% Tween-20.
3. Washing solution: 50 mol/L, pH 7.2 PBS.
4. Specific antibody (primary antibody) or antigen
5. Gold standard antibody
Method of operation
1. Remove the percolator, first add 2 drops (about 100ml) of blocking solution to the small hole NC membrane;
2. After it has penetrated into the box, add 1 drop (about 50 ml) of the specimen to be inspected on the NC membrane;
3. After it has penetrated into the box, add 2 drops (about 100ml) of washing solution on the NC membrane;
4. After it is infiltrated into the box, add 1 drop (about 50 ml) of the gold standard antibody on the NC membrane to react with the adsorbed sample on the NC membrane;
5. After it has penetrated into the box, add 2 drops (about 100 ml) of washing solution on the NC membrane;
6. until the washing liquid completely penetrates into the box
Observation of results
A positive reaction occurred in the red reaction on the small hole NC membrane, and the negative reaction did not stain.
Precautions
1. The specific antibody (primary antibody) or antigen, the sample to be tested and the amount of the gold-labeled antibody must be determined in advance by the square matrix method.
2. The padded material at the bottom of the box should be strong in water absorption, otherwise the result will be affected.
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