Granular tag for biological detection
A biological detection and granular technology, applied in the direction of biological testing, material inspection products, etc., can solve the problems of cumbersome preparation process, multi-labeling, and long time-consuming, etc., and achieve the effect of high adsorption efficiency
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Embodiment 1
[0010] Example 1 Direct method
[0011] 1. Take 10-100ml of 0.01% chloroauric acid aqueous solution, add 1-5ml of 0.1-1% trisodium citrate, heat and boil for 5-50 minutes until the color becomes wine red, and the gold particle solution is obtained.
[0012] 2. Take 10ml of the above gold particle solution, and use 0.1mol / LK 2 CO 3 0.1mol / L HCL to adjust the pH to 6-11.
[0013] 3. Take a certain amount of HIV antigen and alkaline phosphatase solution, add it dropwise to the pH-adjusted solution, stir for 10-30 minutes, then add BSA to a final concentration of 0.1-1%.
[0014] 4. Centrifuge at 20000 rpm for 1 hour at low temperature, and discard the supernatant.
[0015] 5. Resuspend the pellet in a buffer solution containing preservatives and stabilizers, and store it at low temperature for later use.
Embodiment 2
[0016] Embodiment 2 indirect method
[0017] 1. Add 50-100ml of deionized water, 30-90ml of acetone, and 10-30g of styrene into a four-neck flask equipped with a stirrer, condenser tube, feeder and nitrogen gas guide device, and put it in a water bath at a certain temperature for 20- After 50 minutes, 0.2-0.5 g of potassium persulfate was added and reacted for 8-12 hours. The obtained emulsion was separated at a high speed of 25,000 rpm, and the precipitate was washed with buffer solution and then diluted to obtain latex particles.
[0018] 2. Add 5-10ml of 0.05-0.1mol / L n-aminocaproic acid and 10-20ml of 0.05-02mol / L pH7.2 buffer solution to 10ml of the above-mentioned latex particles.
[0019] 3. Take 2ml of the above solution, add 0.5-2ml of streptavidin (5000mg / L) and 5-15mg of carbodiimide, mix well, 37°C for 2 hours, add 0.1-0.5ml of buffer solution of PH8-10, After 30 minutes, centrifuge at 10,000 g for 1 hour, discard the supernatant, and wash the pellet twice with pH...
Embodiment 3
[0024] Example 3 Application of particulate markers in HIV antibody detection
[0025] 1. Take an ELISA plate pre-coated with HIV antigen, add 50ul of the serum to be tested to each well; add 50ul of positive control serum and negative control serum to the positive control well and negative control well respectively, and place at 37°C for 1h.
[0026] 2. After washing the plate 5 times with phosphate buffer containing surfactant, add 50 ul of the granular marker prepared according to Example 1 or Example 2 diluted in a certain proportion to each well, and place at 37° C. for 1 hour.
[0027] 3. After washing the plate 3 times with phosphate buffer solution containing surfactant, add 100ul of chromogen to each well and place at 37°C for 15min.
[0028] 4. After adding 50ul of stop solution to each well, place it on a microplate reader to measure the absorbance value of each well, and judge the result according to the absorbance value of each well.
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