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Antibody conjugation kits and multicolor staining

Advantages of conjugation kits for setting up direct multiplex immunoassays

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:

  1. A comparison between indirect and direct assays
  2. Antibody conjugation kits ,a partner in direct assays

1. Indirect vs direct assays

Indirect vs direct

Legend





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

IndirectAllows signal amplificationNonspecific background may result from the secondary antibody cross-reactivity
Immunoreactivity of the primary antibody is not affected by traditional labeling methodExtra incubation steps are required
The ratio of secondary to primary antibody has to be equal
Time consuming
DirectQuick methodology since only one antibody is usedImmunoreactivity of the primary antibody may be reduced after traditional labeling method
No need for secondary antibodiesLittle 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.

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