Method for preparing double-resistant sandwich method test paper for ancient wool fabric
A technology of detection test paper and double-antibody sandwich, which is applied in the field of detection of traces of ancient wool fabrics, can solve the problems of difficult identification of wool products, and achieve the effects of intuitive results, high sensitivity and strong specificity
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Embodiment 1
[0011] Embodiment 1 adopts the following steps:
[0012] A) prepare gold nanoparticle, adopt the method for sodium citrate reduction chloroauric acid to prepare the gold nanoparticle that particle diameter is 25nm; About 100ml mass concentration is that the chloroauric acid solution water bath of 0.01% is heated to boiling, adds 1.0ml rapidly The mass concentration is 1% sodium citrate solution, boils 7min, and solution presents red, promptly makes gold nano-sol;
[0013] B) Prepare gold-labeled rabbit anti-keratin antibody, use 0.1mol / l K 2 CO 3 Solution Adjust the pH of 100ml of colloidal gold solution to 9.2, add 20 μl of rabbit anti-keratin antibody with stirring, and the antibody concentration is 3.15mg / ml; then add 5ml of polyethylene glycol 20000 solution with a mass concentration of 1%, Stir at room temperature for 5 min, then centrifuge at 9000 r / min for 60 min, discard the supernatant; dissolve the precipitate in 0.01 mol / l pH of 8.2 tris buffer, i.e. Tris-HCl solu...
Embodiment 2
[0016] Embodiment 2 adopts the following steps:
[0017] A) prepare gold nanoparticles, adopt the method for sodium citrate reduction chloroauric acid to prepare the gold nanoparticles that particle diameter is 40nm; Be about to 100ml mass concentration be that the chloroauric acid solution water bath of 0.01% is heated to boiling, add 1.25ml rapidly The mass concentration is 1% sodium citrate solution, boils 8min, and solution presents red, promptly makes gold nano-sol;
[0018] B) Prepare gold-labeled rabbit anti-keratin antibody, use 0.1mol / l K 2 CO 3 Solution Adjust the pH of 100ml of colloidal gold solution to 9.2, add 35μl of rabbit anti-keratin antibody with stirring, and the antibody concentration is 3.15mg / ml; then add 5ml of 5% polyethylene glycol 20000 solution at room temperature Stir for 5min, then centrifuge at 0000r / min for 50min, discard the supernatant; dissolve the precipitate in 0.01mol / l pH of 8.2 tris buffer, i.e. Tris-HCl solution, the Tris -The mass c...
Embodiment 3
[0021] Embodiment 3 adopts the following steps:
[0022] A) prepare gold nanoparticles, adopt the method for sodium citrate reduction chloroauric acid to prepare the gold nanoparticles that particle diameter is 50nm; Be about to 100ml mass concentration be that the chloroauric acid solution water bath of 0.01% is heated to boiling, add 1.5ml rapidly The mass concentration is 1% sodium citrate solution, boils 10min, and solution presents red, promptly makes gold nano-sol;
[0023] B) Prepare gold-labeled rabbit anti-keratin antibody, use 0.1mol / l K 2 CO 3 Solution Adjust the pH of 100ml of colloidal gold solution to 9.2, add 50μl of rabbit anti-keratin antibody with stirring, and the antibody concentration is 3.15mg / ml; then add 5ml of 10% polyethylene glycol 20000 solution at room temperature Stir for 5min, then centrifuge at 11000r / min for 40min, discard the supernatant; dissolve the precipitate in 0.01mol / l pH of 8.2 tris buffer, i.e. Tris-HCl solution, the Tris -The mass...
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