Thyroid microsomal antibody detection reagent kit as well as preparation method and application thereof
A technology of thyroid microsomes and kits, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of rare detection kits, and achieve the effects of long detection time, long storage time, and small system error
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[0049] Example 1
[0050] (1) Labeling of protein A
[0051] Preparation of dialysate (F solution): add Na in a 5000ml beaker 2 CO 3 14.31g, NaHCO 3 26.46g, diluted with purified water to 4500ml. The prepared F solution was placed on a magnetic stirrer for use.
[0052] Select a dialysis bag with a suitable retention volume (usually 14000), measure the appropriate size, tie one end tightly after wetting, and test the purified water for leakage 3 times (no leakage is required).
[0053] Take 100 μg of protein A and adjust it to 1 ml with 0.1 mol / L pH9.5 carbonate buffer (F solution). Put it into the dialysate, stir and dialyze at room temperature for 2 hours, add 300 μg of ABEI activated ester to the dialyzed solution, and react at 37° C. for 2 hours.
[0054] The ligation product of protein A and ABEI was purified by G-25 gel column.
[0055] D 2 Preparation of solution: add 200ml of 0.5M phosphate buffer (P001 solution), 20g BSA, 8g NaN 3 , 2g MgCl 2 ·6H 2 O, 600ml...
Example Embodiment
[0072] Example 2
[0073] (1) Protein A biotinylation, the specific steps are as follows:
[0074] Preparation of dialysate (F solution): add Na in a 5000ml beaker 2 CO 3 14.31g, NaHCO 3 26.46g, diluted with purified water to 4500ml. The prepared F solution was placed on a magnetic stirrer for use.
[0075] 100 μg of biotin and 1 mg of protein A were taken and adjusted to 1 ml with 0.1 mol / L pH9.5 carbonate buffer (F solution). Put it into the dialysate, stir and dialyze at room temperature for 2 hours, and react at 37°C for 2 hours.
[0076] Purified by G-25 gel column.
[0077] The D2 solution was prepared according to the method of Example 1, and the purified ligation product was D2 2 The solution was diluted to a concentration of 0.025 μg / ml.
[0078] (2) Marking of SA
[0079] 100 μg of SA was adjusted to 1 ml with 0.1 mol / L carbonate buffer (F solution) of pH 9.5. Put it into the dialysate, stir and dialyze at room temperature for 2 hours, add 300 μg of ABEI ac...
Example Embodiment
[0095] Example 3
[0096] (1) Labeling of protein A
[0097] Preparation of dialysate (F solution): add Na in a 5000ml beaker 2 CO 3 14.31g, NaHCO 3 26.46g, diluted with purified water to 4500ml. The prepared F solution was placed on a magnetic stirrer for use.
[0098] Select a dialysis bag with a suitable retention volume (usually 14000), measure the appropriate size, tie one end tightly after wetting, and test the purified water for leakage 3 times (no leakage is required).
[0099] Take 100 μg of protein A and adjust it to 1 ml with 0.1 mol / L pH9.5 carbonate buffer (F solution). Put it into the dialysate, stir and dialyze at room temperature for 2 hours, add 300 μg of ABEI activated ester to the dialyzed solution, and react at 37° C. for 2 hours.
[0100] The ligation product of protein A and ABEI was purified by G-25 gel column.
[0101] D 2 Preparation of solution: add 200ml of 0.5M phosphate buffer (P001 solution), 20g BSA, 8g NaN 3 , 2g MgCl 2 ·6H 2 O, 600ml...
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