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Coupling antibody to cyanogen bromide-activated sepharose.
Jennifer M Kavran1, Daniel J Leahy1
1Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Methods in Enzymology
|March 29, 2014
Summary
Antibodies are immobilized on cyanogen bromide-activated Sepharose. This method enables their use in pull-down assays and immunoaffinity purification for biological research.
Area of Science:
- Biochemistry
- Immunology
- Protein Chemistry
Background:
- Antibody immobilization is crucial for various biochemical assays.
- Efficient methods are needed for antibody-based purification and detection.
Purpose of the Study:
- To describe a method for immobilizing antibodies onto Sepharose beads.
- To enable the use of immobilized antibodies in pull-down assays and immunoaffinity purification.
Main Methods:
- Antibodies are covalently attached to cyanogen bromide-activated Sepharose beads.
- The immobilized antibodies are prepared for subsequent experimental procedures.
Main Results:
- Successful immobilization of antibodies on Sepharose.
- Prepared beads are ready for pull-down assays and immunoaffinity purification.
Conclusions:
- Cyanogen bromide-activated Sepharose provides a reliable matrix for antibody immobilization.
- This technique supports robust pull-down assays and immunoaffinity purification protocols.
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