Published: Vol 2, Iss 6, Mar 20, 2012 DOI: 10.21769/BioProtoc.133 Views: 22957
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Abstract
This protocol describes a method of purifying antibodies from sera with denatured antigens immobilized on western blot membranes. Advantages include (1) fast and easy; (2) purification of antibody with antigen in denatured form allows high yield in case antigen protein solubility is limited. Disadvantage is that possible antibodies that recognize certain 3D structure in solution of antigens might not be purified using such a method. Regarding this issue, the flow through is recommended to be kept and can be used for other purification methods with folded antigens.
Materials and Reagents
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Recipes
Acknowledgments
This protocol was adapted from Kurien (2009) and developed in Guowei Fang’s lab, Departmental of Biological Sciences, Stanford University, CA, USA. Funding from the NIH supported this work.
References
Article Information
Copyright
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Fang, L. (2012). Antibody Purification from Western Blotting. Bio-protocol 2(6): e133. DOI: 10.21769/BioProtoc.133.
Category
Immunology > Antibody analysis > Antibody-antigen interaction
Immunology > Antibody analysis > Antibody detection
Biochemistry > Protein > Immunodetection
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