Unlock instant, AI-driven research and patent intelligence for your innovation.

Production method and application method of closing-free ELISA (Enzyme Linked Immunosorbent Assay) plate

A production method and a technology for an ELISA plate, which are applied in the field of analysis and detection, can solve the problems of high noise, waste users, and take a long time, and achieve the effects of reliable accuracy, time saving and easy operation.

Active Publication Date: 2019-01-01
SHENZHEN UNIV
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the above step (2) is essential, because without blocking, it will lead to non-specific adsorption of the subsequent enzyme-labeled secondary antibody, that is, samples without the antigen or antibody to be detected will also have signals and high noise, which will affect the detection Sensitivity
However, closing this step usually takes the user nearly two hours, and the steps are cumbersome and time-consuming

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Production method and application method of closing-free ELISA (Enzyme Linked Immunosorbent Assay) plate
  • Production method and application method of closing-free ELISA (Enzyme Linked Immunosorbent Assay) plate
  • Production method and application method of closing-free ELISA (Enzyme Linked Immunosorbent Assay) plate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] This specific embodiment 1 provides a kind of manufacture method of free-blocking ELISA microplate, and among the present invention, microplate uses 96 orifice plates, and the steps are as follows:

[0025] Step 1. Prepare sodium citrate colloidal gold solution; soak the round-bottomed flask used for synthesizing sodium citrate colloidal gold with aqua regia for 24 hours, and wash it with deionized water for later use. Add 1000mL deionized water and 12mL 0.01g / mL trisodium citrate solution to the above-mentioned round bottom flask, and boil; then add 10mL 0.01g / mL chloroauric acid, continue to boil for 15min, and cool naturally to obtain a concentration of 40μg / mL sodium citrate colloidal gold solution, and place it in a refrigerator at 4°C for later use.

[0026] Step 2. Add 100 μL of PEI (polyethyleneimine) aqueous solution with a concentration of 5 mg / mL into the experimental wells of a 96-well plate (common microplate plate without sealing), soak for 2 hours, disca...

Embodiment 2

[0031] The specific embodiment 2 is basically the same as the embodiment 1, except that the concentration of the PEI aqueous solution in step 2 is 0.1 mg / mL.

Embodiment 3

[0033] This specific example 3 is basically the same as Example 1, except that the concentration of the sodium citrate colloidal gold solution in step 3 is 10 μg / mL.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
concentrationaaaaaaaaaa
Login to View More

Abstract

The invention provides a production method of a closing-free ELISA (Enzyme Linked Immunosorbent Assay) plate. The production method comprises the following steps: step I, separately adding a polyethyleneimine aqueous solution with the concentration of 0.1mg / mL to 5mg / mL into experiment holes of the ELISA plate, soaking, removing residual solution, and cleaning by using deionized water; step II, adding a sodium citrate colloidal gold solution with the concentration of 10 microg / mL to 40 microg / mL into the experiment holes of the ELISA plate in step I, soaking, removing the residual solution, cleaning by using deionized water to obtain the polyethyleneimine-gold nano particle-modified ELISA plate; and step III, drying, sealing and packaging the ELISA plate obtained in step II, thus obtainingthe finished product plate. By adopting the production method of the closing-free ELISA plate, the closing step in the ELISA process can be saved, and the protein detection time is shortened on the premise of not influencing the detection sensitivity and specificity.

Description

technical field [0001] The invention belongs to the technical field of analysis and detection, and relates to a method for making and using a non-blocking ELISA plate. Background technique [0002] Enzyme Linked Immunosorbent Assay (ELISA) is a detection reaction based on the specific interaction between antigen and antibody. The detection of targets by ELISA is mainly carried out by the double-antibody sandwich method or the double-antigen sandwich method, which mainly includes the following steps: (1) immobilizing the capture antibody or antigen on a solid-phase substrate (enzyme plate); (2) using bovine serum albumin (3) Antibody or antigen incubation reaction; (4) Enzyme-labeled secondary antibody incubation reaction; (5) Determination of color development, luminescence or gas production by detecting the enzyme molecule labeled on the antibody to catalyze the substrate target concentration. Among them, the above step (2) is essential, because without blocking, it will ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCG01N33/54346
Inventor 陈雯雯张克黄瑞嘉
Owner SHENZHEN UNIV