A kind of preparation method of elisa pre-coated microtiter plate
A technology of microplate plate and pre-coating, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of insufficient storage time and scrapped ELISA kits, so as to increase storage time and stability, reduce economic losses, The effect of reducing scrap
Active Publication Date: 2016-04-13
杭州联科生物技术股份有限公司
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Problems solved by technology
In recent years, there has been a blocking step after the coating step, and drying after sealing; but at present, there is no fixative specially used for immobilizing protein activity between the blocking and drying steps, so that the storage time is not long enough, resulting in the failure of ELISA kits. The pre-coated ELISA plate expired and was scrapped, resulting in considerable economic losses
Method used
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Experimental program
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Effect test
Embodiment 1
[0016] Example 1: Preparation steps of HumanIL-6 pre-coated microtiter plate:
[0017] a. Preparation of coating solution: Dilute the anti-HumanIL-6 capture antibody to 2ug / ml with PBS buffer solution with a phosphate molar concentration of 0.01mol / L and pH=7.2 to obtain a coating solution;
[0018] b. Coating: Add 100ul of coating solution to the enzyme-labeled well, and coat overnight at 4°C;
[0019] c. Blocking: Discard the coating solution, add 300ul / well of 1%BSA-PBST blocking solution with pH=7.3, and block at room temperature for 4 hours;
[0020] d. Preparation of fixative solution: add sucrose and trehalose to PBS buffer solution with phosphate molar concentration of 0.01mol / L and pH=7.3, so that the mass fraction of sucrose is 30%, and the mass fraction of trehalose is 30%. Get the fixative.
[0021] e. Fixation: Discard the blocking solution, add 300ul of the above fixing solution per well, and fix at room temperature for 0.5 hours;
[0022] f. Drying: Discard t...
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The invention relates to a preparation method for a pre-coated enzyme-linked immunosorbent assay (ELISA) plate and belongs to the technical field of diagnostic reagents. The preparation method includes two steps of preparing coating solution and coating, and further includes the following steps: a, confining: discarding the coating solution, adding confining solution to each hole with 100-300 microlitres, and then carrying out confining at room temperature for four hours; b, preparing fixing : adding sucrose and trehalose to a phosphate buffer solution (PBS) with the phosphate radical molarity of 0.01mol / L and the potential of hydrogen (pH) of 7.2-7.4 to enable the mass fraction of the sucrose to be 5-30% and the mass fraction of the trehalose to be 5-30%, and then obtaining the fixing solution; c: fixing: discarding the confining solution, adding the fixing solution to each hole with 100-300 microlitres, and fixing the solution for 0.5 hour; and d, drying: discarding the fixing solution, drying in the air at room temperature for a night, evacuating an aluminium foil bag to a vacuum state, adding a drying agent and preserving the aluminium foil bag at 4 DEG C to obtain the pre-coated ELISA plate. The pre-coated ELISA plate prepared with the method can be preserved at 4 DEG C for more than three years.
Description
technical field [0001] The invention relates to a preparation method of an ELISA pre-coated microplate, and belongs to the technical field of biological in vitro diagnostic reagents. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA) has become an internationally recognized gold standard for protein quantification due to its high sensitivity, easy access to samples, and simple operation. One of the key technologies in the preparation of ELISA kits is the preparation of antibody solid-phase carrier microtiter plates. The conventional preparation method of the pre-coated ELISA plate is to incubate the coating solution in the enzyme-labeled well for a certain period of time, then discard the coating solution, store it at 4°C for 1 month, and store it at -20°C for 3 months; After adding desiccant and vacuuming, it can be stored at 4°C for about 1 year, but this cannot meet the needs of long-term storage of ELISA kits. ELISA pre-coated microtiter plates requ...
Claims
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IPC IPC(8): G01N33/543
Inventor 袁志波
Owner 杭州联科生物技术股份有限公司