Antibody conjugation kits help you label your primary antibody library. By conjugating your primary antibodies, you can set up a direct assay and avoid potential cross-reactivity of the secondary antibodies.
In contrast to traditional labeling methods, the conjugation process with our conjugation kits does not affect the immunoreactivity of your primary antibody.
Discover below:
- A comparison between indirect and direct assays
- Antibody conjugation kits ,a partner in direct assays
1. Indirect vs direct assays
Figure: Indirect versus direct assays
The indirect (two-step) method uses a labeled secondary antibody for detection. First, a primary antibody is incubated with the antigen. This is followed by incubation with a labeled secondary antibody that recognizes the primary antibody.
In contrast, the direct assay uses the method of directly labeling the antibody itself. The antibody is conjugated with a colorimetric, chemiluminescent or fluorescent label. Since the secondary antibody step is omitted, the direct assay is relatively quick and avoids potential problems of cross-reactivity of the secondary antibody with components in the antigen sample.
Table: Comparison between indirect and direct assays
Method | Advantages | Disadvantages |
|---|---|---|
| Indirect | Allows signal amplification | Nonspecific background may result from the secondary antibody cross-reactivity |
| Immunoreactivity of the primary antibody is not affected by traditional labeling method | Extra incubation steps are required | |
| The ratio of secondary to primary antibody has to be equal | ||
| Time consuming | ||
| Direct | Quick methodology since only one antibody is used | Immunoreactivity of the primary antibody may be reduced after traditional labeling method |
| No need for secondary antibodies | Little signal amplification | |
| No cross-reactivity of secondary antibody | ||
| Same primary antibody species can be used in the same experiment | ||
| Different fluorochromes can be used in the same experiment |
2. Antibody conjugation kits, a partner in direct assays
In contrast to traditional labeling methods, our antibody conjugation process does not affect the immunoreactivity of your primary antibody.
Antibody conjugation kits eliminate:
- Indirect detection methods
- Column separation steps
- Loss of material
- Non-specific binding of secondary reagents
- Additional incubation steps and sample dilution
We offer a large range of labels suitable for enzyme, biotin/streptavidin and fluorochrome detection methods.
- Enzymatic labels: AP, Glucose oxidase and HRP
- Precipitating agents: Avidin, Biotin and Streptavidin
- Fluorochromes: AMCA, Cy® dyes, FITC, PE, PerCP, Rhodamine and Texas Red®
- Fluorescent tandem dyes: APC, PE or PerCP combined with Cy® dyes. Learn more about tandem dyes.
View our resources and FAQs to find the right antibody conjugation kit.