| Chapter title |
Affinity Purification of Antibodies
|
|---|---|
| Chapter number | 3 |
| Book title |
ELISA
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-2742-5_3 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-2741-8, 978-1-4939-2742-5
|
| Authors |
Robert M. Hnasko, Jeffery A. McGarvey, Hnasko, Robert M., McGarvey, Jeffery A. |
| Abstract |
Antibodies are provided in a variety of formats that include antiserum, hybridoma culture supernatant, or ascites. They can all be used successfully in crude form for the detection of target antigens by immunoassay. However, it is advantageous to use purified antibody in defined quantity to facilitate assay reproducibility, economy, and reduced interference of nonspecific components as well as improved storage, stability, and bio-conjugation. Although not always necessary, the relative simplicity of antibody purification using commercially available protein-A, protein-G, or protein-L resins with basic chromatographic principles warrants purification when antibody source material is available in sufficient quantity. Here, we define three simple methods using immobilized (1) protein-A, (2) protein-G, and (3) protein-L agarose beads to yield highly purified antibody. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 27 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Doctoral Student | 5 | 19% |
| Student > Ph. D. Student | 5 | 19% |
| Researcher | 4 | 15% |
| Student > Bachelor | 3 | 11% |
| Other | 2 | 7% |
| Other | 3 | 11% |
| Unknown | 5 | 19% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 10 | 37% |
| Agricultural and Biological Sciences | 5 | 19% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 4% |
| Immunology and Microbiology | 1 | 4% |
| Physics and Astronomy | 1 | 4% |
| Other | 1 | 4% |
| Unknown | 8 | 30% |