Technical Information on Biotin Antibody Conjugates


The high affinity binding of avidin with biotin provides a unique approach to antibody localization. PROTOS IMMUNORESEARCH supplies affinity purified antibodies and F(ab')2 or Fab' antibody fragments conjugated to biotin. These reagents can be localized by utilizing (strept)avidin conjugates that specifically bind biotin. Experience has shown that biotin conjugates are sensitive and specific tools in immunohistology, immunocytology, ELISA and immunoblotting. The working dilution of biotin conjugated antibodies in ELISA is 1:10,000 - 1:20,000.

Introduction of Biotin groups in Monoclonal Antibodies

Biotin conjugated goat Fab'anti-mouse IgG(H+L) [Cat.No: 648] can be used to introduce biotin groups into mouse monoclonals:

  1. Dilute the monoclonal antibody to 200 ng/ml in PBS. (1:5000 when starting from 1 mg/ml).
  2. Add Cat.648 to a final conc. of 1:1000. Mix well.
  3. Incubate 30 minutes at room temperature.
  4. Bind excess Cat.648 by adding 1:100 final normal mouse serum. Mix well.
  5. Incubate 30 minutes at room temperature.

Since excess Cat.648 has been removed, the now biotin conjugated monoclonalantibody can be used on a variey of proteins (eg Western Blots) and tissues, including mouse.

In a similar fashion, biotin conjugated goat Fab' anti-rat IgG(H+L) [Cat.No: 687] can be used to introduce biotin groups into rat monoclonals. Excess Cat.687 will be removed with 1:100 final normal rat serum.          


 

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875A Cowan Road, Burlingame, CA 94010
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