ELISA testing and Kits development services丨Zoonbio

Summary

ELISA (Enzyme-linked Immunosorbent Reaction Analysis) is a highly sensitive experimental technique which combines the specific reactions of antigens and antibodies with the efficient catalysis of enzymes on substrates based on immunological reactions. Because the reaction of antigen and antibody is carried out in the pore of polystyrene microtitration plate as a solid carrier, after incubation with each reagent, the excess free reactants can be removed by washing, thus ensuring the specificity and stability of the test results.

Zoonbio, together with the well-known foreign trade ELISA kit brand in China, has launched more than 20,000 ELISA detection kits. All customers who buy the kits in Zoonbio can enjoy free ELISA detection services.

ELISA testing experiment steps

ELISA(detection of unknown antigens)	indirect method(detection of unknown antibodies)
1.The antibody was diluted to 1-10 ug/ml with buffer.0.1 ml was added into the reaction pore and spent the night at 4 C.The next day, the solution in the hole was discarded and washed three times with a washing buffer.	1.The known antigens were diluted to 1-10 ug/ml with coated buffer, 0.1 ml per hole, and overnight at 4 C. Wash three times the next day.
2.Sample addition:A diluted sample of 0.1 ml was incubated in the coated reaction pore at 37 C for 1 hour.Then wash (blank pore, negative control pore and positive control pore were made at the same time).	2.Sample addition:A diluted sample (unknown antibody) of 0.1 ml was incubated in the coated reaction pore at 37 C for 1 hour and washed. (blank control, negative control and positive control were also performed).
3.Enzyme-labeled antibodies:Fresh diluted enzyme-labeled antibodies (dilution after titration) were added to each reaction pore at 0.1 ml. Incubate at 37 C for 0.5-1 hour and wash.	3.Enzyme-labeled antibodies:In the reaction pore, fresh diluted enzyme-labeled second antibody (anti-antibody) 0.1ml was added, incubated at 37 C for 30-60 minutes, washed, and washed with DDW for the last time.
4.Adding Substrate Solution to Colour Development:Temporarily prepared TMB substrate solution was added to each reaction pore at 0.1ml, 37 C for 10-30 minutes.	4.Adding Substrate Solution to Colour Development:Temporarily prepared TMB substrate solution was added to each reaction pore at 0.1ml, 37 C for 10-30 minutes.
5.Termination reaction:2 M sulphuric acid 0.05 ml was added into each reaction pore.	5.Termination reaction:2 M sulphuric acid 0.05 ml was added into each reaction pore.
6.Result judgement:The results can be observed directly with naked eyes on a white background.The darker the color in the reaction pore, the stronger the positive degree. The negative reaction is colorless or very light. According to the depth of the color, it is indicated by "+" and "-".OD value can also be measured: On ELISA detector, at 450 nm (410 nm if ABTS is used to color), OD value of each hole is measured after zero adjustment of blank control hole. If OD value of each hole is greater than 2.1 times of the prescribed negative control OD value, it will be positive.	6.Result judgement:The results can be observed directly with naked eyes on a white background.The darker the color in the reaction pore, the stronger the positive degree. The negative reaction is colorless or very light. According to the depth of the color, it is indicated by "+" and "-".OD value can also be measured: On ELISA detector, at 450 nm (410 nm if ABTS is used to color), OD value of each hole is measured after zero adjustment of blank control hole. If OD value of each hole is greater than 2.1 times of the prescribed negative control OD value, it will be positive.

ELISA testing service process

1.If there is any need, contact us by telephone, mail, fax or business personnel, our technical support staff will communicate with you in time, confirm the order, and negotiate the experimental scheme. After signing the customization agreement, 50% of the total price will be paid as advance payment.

2.According to the requirements of the experiment, the necessary antibodies and antigens are provided. It is generally believed that the commercial antibodies meet the requirements of ELISA experiment. When customers provide antigen-conjugated antibodies, please provide negative control to ensure the accuracy and accuracy of the experiment.

3.During the experimental stage, our technical support staff will report the progress of the experiment to you in time.

4.At the end of the experiment, according to the contract signed, the remaining experimental materials will be returned and the test report will be sent to you. At the same time, you will be asked to pay the remaining 50% of the payment.

Development of ELISA Kit

We adhere to the eight quality control principles of ELISA kit development and successfully develop highly sensitive, economical and practical disease detection kits for aquatic and animal disease detection users.

1 principle of stability Ambient temperature for 2 weeks	2 principle of applicability detectable natural samples
3 recovery Principle recovery control 75%-125%	4 cross reaction
5 precision Principle the difference between plates and between plates was less than 15%	6 minimum detection limit depending on the actual testing environment
7 linear relationship	8 specificity

STEP	DETAIL
antibody labeling and pairing	The purified monoclonal/polyclonal antibodies of X protein were labeled with HRP to determine whether the antibody pairs of sandwich positive samples could be formed.
optimization of working concentration	The sensitivity and specificity of antigen-antibody reaction were optimized by concentration. Determining the optimal concentration of antigen and antibody.
optimization of coating conditions and reaction conditions	The coating buffer, washing buffer and preservation solution of enzyme conjugate were determined, and the reaction temperature and time of antibody were determined.
measurement of inter-batch difference and inter-batch difference	Repeatability test, inter-batch difference and inter-batch difference should be less than 15%.
stability test	The performance of the kit was tested by 37 degree destructive test for 3 days and 7 days at 37 degree. At the same time, the samples are kept at 4 degrees for long-term detection. The general kit can be stored at 4 degrees for more than half a year.