Development of Indirect ELISA
Creative Diagnostics has advanced scientific research concepts, strong development strength, and has rich experience in various immunoassays. We offer our customers thousands of reliable, ready-to-use indirect ELISA kits. On this basis, we provide indirect ELISA development services for the special needs of customers. We are very confident that this service can fully meet the needs of customers and develop new ELISA detection methods for customers.
Why Choose Indirect ELISA Development?
When you need to detect an antibody and can't find an existing ELISA kit when you expect to detect the antibody at a lower concentration but can't find an ELISA kit that meets the requirements, or you are developing a new indirect ELISA detection method for this antibody, at this time, you can choose us to carry out the development of indirect ELISA for you with confidence.
Questions to Consider Before Choosing This Service
- Is your analyte an antibody?
- Do you need to amplify the detection signal?
- Are there multiple suitable antibodies for your target antigen?
If your answer is yes, this is a preliminary indication that indirect ELISA can meet your development needs. If you need further confirmation, please contact me directly. Our professional team will answer you whether this service can meet your requirements.
Advantages and Disadvantages of Indirect ELISA
In an indirect ELISA, the antigen is first bound to a microplate. After washing to remove excess antigen, the sample to be tested is added, and the antibody in the sample specifically binds to the antigen, and then an enzyme-labeled secondary antibody is added for color development and determination.
The Main Advantages of Indirect ELISA
- The use of enzyme-labeled secondary antibodies amplifies the reaction signal, making it more sensitive.
- Different primary antibodies can be used with a single labeled secondary antibody, making it more flexible.
The Main Disadvantages of Indirect ELISA
- The enzyme-labeled secondary antibody binds directly to the antigen and may cross-react.
- The experiment added the process of secondary antibody incubation, which extended the experimental period.
Fig. 1 The Flowchart of Indirect ELISA
Development of Indirect ELISA
The development of indirect ELISA includes the steps of formulating a protocol, preparing antigens, preparing antibodies, preparing enzyme-labeled antibodies, and selecting ELISA components. To develop accurate ELISA kits for customers, we strictly implement the following indirect ELISA development service process.
Our professional team will analyze your testing needs and determine that there is no indirect ELISA kit on the market that meets your needs. We will design an indirect ELISA development plan that fully meets your requirements from sensitivity to specificity.
In terms of the development of indirect ELISA, the preparation of the required antigen is a crucial step. With our solid expertise, we prepare high-purity antigens in expression systems including E. coli and yeast.
Based on the prepared antigens, we use our antibody preparation development platform to prepare primary and secondary antibodies that meet experimental needs.
After synthesizing the desired antibody, we analyze the level of biomolecular interactions using antibody affinity assays to ensure specific binding between the antigen and antibody.
Our expert team will select the most appropriate method to label the secondary antibody among radioisotope labeling, biotin labeling, fluorescent labeling, etc.
First, we will select various reagents used in indirect ELISA (such as coating buffer, sample diluents, assay diluents, conjugate stabilizers, stop solution, PBS buffer, etc.) and various components (ELISA Plate, Foil bags, desiccant packets, etc.). The indirect ELISA protocol development was then started.
In this step, we will optimize the sandwich ELISA for issues such as non-specific binding of antibodies, high substrate concentration, poor reproducibility, etc.
When the indirect ELISA development protocol is optimized, in order to ensure that the protocol is completely accurate, we also perform assay validation on your specified samples.
After verifying that the indirect ELISA development plan is correct, we will produce indirect ELISA kits according to your requirements, and provide pre-coated plates and all reagents.
Result/Material Delivery
Development Protocol
All Experi-mental Data
Ready-to-use ELISA Kits
Validated Assay Results
The Creative Diagnostics team has many years of experience in ELISA development, and all services are provided according to your individual needs. Please contact us for more information on our development of indirect ELISA. At the same time, we also provide ELISA testing services and provide comprehensive solutions for your ELISA kit customization needs.
