Method for preparing streptavidin pre-coated elisa plate and application

A streptavidin and pre-coating technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome process, unstable streptavidin fixation, and no streptavidin yet. The preparation method is simple, suitable for long-term storage, and the effect of coating is excellent.

Inactive Publication Date: 2010-04-07
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The wet coating method optimizes the coating effect by adjusting the concentration of streptavidin and the coating buffer. The process is cumbersome and takes a long time. Low, the coated streptavidin is unstable, and it is easy to wash off during plate washing
Moreover, the wet coating method has high requirements on the adsorption force of the microtiter plate, and most of the coating solution must be discarded after the coating is completed, the utilization rate of streptavidin is low, and the cost is high
The dry coating method is rarely used in ELISA coating because it will affect the activity and spatial structure of antigens and antibodies
So far, there is no report on streptavidin pre-coated microtiter plate prepared by dry coating method

Method used

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  • Method for preparing streptavidin pre-coated elisa plate and application
  • Method for preparing streptavidin pre-coated elisa plate and application
  • Method for preparing streptavidin pre-coated elisa plate and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The blank microtiter plate is Jiete high affinity microtiter plate.

[0027] Use pH 9.6, 0.05M carbonate buffer (1.59g sodium carbonate and 2.93g sodium bicarbonate diluted with water to 1000mL) to dilute streptavidin to 2 μg / ml to obtain coating buffer; Add it to a blank microtiter plate, 100 μl per well, and then incubate overnight at 37°C in an incubator until completely dry to obtain a streptavidin pre-coated microtiter plate. The dried microtiter plates should be packed in sealed bags and stored at 4°C.

Embodiment 2

[0029] The blank microtiter plate is Jiete high affinity microtiter plate.

[0030] Use pH 5.0, 0.15M citrate phosphate buffer (1030ml 0.1M citric acid and 970ml 0.2M Na 2 HPO 4 Mix to obtain) dilute streptavidin to 2 μg / ml to obtain coating buffer; add coating buffer to a blank microplate plate, 100 μl per well, and then incubate overnight at 37°C in an incubator until complete Dry to obtain a streptavidin pre-coated microtiter plate. The dried microtiter plates should be packed in sealed bags and stored at 4°C.

Embodiment 3

[0032] Blank microtiter plate is Nunc microtiter plate.

[0033] Use pH 5.0, 0.15M citrate phosphate buffer (1030ml 0.1M citric acid and 970ml 0.2M Na 2 HPO 4 Mix to obtain) dilute streptavidin to 2 μg / ml to obtain coating buffer; add coating buffer to a blank microplate plate, 100 μl per well, and then incubate overnight at 37°C in an incubator until complete Dry to obtain a streptavidin pre-coated microtiter plate. The dried microtiter plates should be packed in sealed bags and stored at 4°C.

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Abstract

The invention discloses a streptavidin pre-coated elisa plate, a preparation method thereof and application thereof. The preparation method comprises the following steps of: dissolving streptavidin by using carbonate buffer solution or citric acid phosphate buffer solution to obtain coated buffer solution; and adding the coated buffer solution into an elisa plate, and then drying the elisa plate to obtain the streptavidin pre-coated elisa plate. The preparation method is simple, and the utilization rate of the streptavidin is high so as to reduce the dosage of the streptavidin. The preparation method has low requirement for the quality of a blank elisa plate; and the batch of the obtained streptavidin pre-coated elisa plate is stable, and the hole difference, batch difference and stability of the streptavidin pre-coated elisa plate are higher than those of wet coating, so the streptavidin pre-coated elisa plate is more suitable for long-term storage. The streptavidin pre-coated elisa plate has low cost and good effect, and can be applied in the technical field of in vitro immune detection or the field of nucleic acid detection in molecular biology.

Description

technical field [0001] The invention belongs to the technical field of in vitro immunoassay, and in particular relates to a streptavidin pre-coated microplate plate and its preparation method and application. Background technique [0002] Streptavidin is a basic glycoprotein secreted by Streptomyces, which is easy to combine with polystyrene plastic microwell plates. One streptavidin molecule can bind to 4 biotin molecules, and the affinity constant (K) of the two is 10 15 L / mol. Due to the extremely high binding force between biotin and avidin, the biotin-avidin system (biotin-avidin system, BAS) has been widely used in ELISA. [0003] The method of immobilizing streptavidin on a solid phase carrier and using BAS to immobilize difficult-to-coat small molecule antigens, antibodies or nucleic acids has been widely used. Streptavidin pre-coated ELISA plates are widely used in ELISA, PCR-ELISA and other fields. The wide application of BAS immobilization depends on the good ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 吴希阳刘宾
Owner JINAN UNIVERSITY
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