Quantitative measurement kit for glypican and detection method thereof
A technology of phosphatidylinositol and proteoglycan, which is applied in the quantitative detection kit of phosphatidylinositol and proteoglycan, can solve the problems of limiting sensitivity and specificity, and achieve the expansion of detection range, large binding capacity, The effect of shortening the reaction time
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[0038] The preparation method of the quantitative assay kit of Glypican (GPC3) of the present invention comprises the following steps:
[0039] The first step: the preparation process of magnetic separation reagent
[0040] 1. Operation steps for preparing magnetic bead buffer, prepare 1L:
[0041] 1. Weigh TRIS 6.06g, NaCl 9.0g, 1.0g TWEEN-20 in a 1L container;
[0042] 2. Use a pipette to measure 0.5ml of Proclin-300 into the above 1L container, then add 800ml of purified water to the above 1L container, and stir thoroughly;
[0043] 3. Adjust the pH meter to measure the pH value, adjust the pH, and control the pH at 8.0 (plus or minus 0.5);
[0044] 4. Weigh 2g of BSA and pour it into the above 1L container;
[0045] 5. Finally, adjust the volume to 1000ml, and filter with a 0.2um filter; after filtering, label it and store it in a cold room at 2-8°C.
[0046] 2. Preparation process of magnetic separation reagent
[0047] 1. Dissolve 2.0mg of disuccinimidyl suberate in...
Embodiment 1
[0087] The magnetic microspheres containing the anti-GPC3 monoclonal antibody of this embodiment were prepared according to the following steps:
[0088] 1. The configuration of the magnetic bead buffer: the formula is shown in Table 1, taking the preparation of 1L as an example:
[0089] 1. Weigh TRIS 6.06g, NaCl 9.0g, 1.0g TWEEN-20 in a 1L container;
[0090] 2. Use a pipette to measure 0.5ml of Proclin-300 into the above 1L container, then add 800ml of purified water to the above 1L container, and stir thoroughly;
[0091] 3. Adjust the pH meter to measure the pH value, adjust the pH, and control the pH at 8.0+0.05;
[0092] 4. Weigh 2g of BSA and pour it into the above 1L container;
[0093] 5. Finally, adjust the volume to 1000ml, and filter with a 0.2um filter; after filtering, label it and store it in a cold room at 2-8°C.
[0094] The magnetic bead buffer is valid for 12 months;
[0095] Table 1 Raw materials and proportioning of magnetic bead buffer
[0096]
[...
Embodiment 2
[0110] The alkaline phosphatase-labeled anti-GPC3 monoclonal antibody of this embodiment was prepared according to the following steps:
[0111] 1. Preparation of enzyme reactant (alkaline phosphatase-labeled anti-GPC3 monoclonal antibody) diluent: the formula is shown in Table 2, taking the preparation of 1L as an example:
[0112] 1. Put 6.06g of Tris, 9.0g of NaCl, 0.05g of Zncl2, 0.5ml of Proclin-300 and 0.05g of MgCl2 in a flask, then add 800ml of purified water into the flask, and stir well to completely dissolve the reagents;
[0113] 2. Adjust PH, control PH in the range of 7.35-7.45;
[0114] 3. Weigh 2g of BSA and pour it into the above beaker;
[0115] 4. Finally, adjust the volume to 1000ml, filter it with a 0.2um filter, label it and store it in a cold room at 2-8°C. The validity period is 12 months.
[0116] Raw materials and proportioning of table 2 enzyme reactant dilution
[0117]
[0118] 2. Coupling of alkaline phosphatase (ALP) and anti-GPC3 monoclona...
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