|
Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences >
Peer-reviewed Journal Articles, etc >
Non-specific adsorption of fish immunoglobulin M (IgM) to blocking reagents on ELISA plate wells
| Title: | Non-specific adsorption of fish immunoglobulin M (IgM) to blocking reagents on ELISA plate wells |
| Authors: | Kim, W.S. Browse this author | | Nishizawa, T. Browse this author | | Yoshimizu, M. Browse this author →KAKEN DB |
| Keywords: | Fish serum | | Immunoglobulin M | | IgM | | Adsorption | | Antibody detection | | Enzymelinked immunosorbent assay | | ELISA |
| Issue Date: | 31-Oct-2007 |
| Publisher: | Inter-Research |
| Journal Title: | Diseases of Aquatic Organisms |
| Volume: | 78 |
| Issue: | 1 |
| Start Page: | 55 |
| End Page: | 59 |
| Publisher DOI: | 10.3354/dao01843 |
| PMID: | 18159673 |
| Abstract: | Enzyme-linked immunosorbent assay (ELISA) is a popular technique for quantifiable detection of specific antibodies in warm-blooded animals, but it has not been accepted for detection of fish antibodies because of its low reproducibility, which is due in part to high background optical density (OD) measurements. In the present study, we report that the high background of a fish antibody- detection ELISA resulted from non-specific adsorption of fish immunoglobulin M (IgM) to blocking reagents on the ELISA plate wells. Four fish sera (from rainbow trout Oncorhynchus mykiss, masu salmon O. masou, Japanese flounder Paralichthys olivaceus and koi Cyprinus carpio) were poured into ELISA plate wells pre-blocked with several blocking reagents (skim milk, soybean milk, bovine serum albumin, fetal bovine serum, gelatin and Roche BlockingReagent®) and then washed out in order to measure the remaining fish IgM on the ELISA plate wells. Significant amounts of fish IgMs (OD absorbance at 492 nm: 0.3 to 1.1) remained on the ELISA plate wells with no antigenic protein except blocking reagents. The amount of remaining fish IgMs on the ELISA plate wells decreased significantly following treatment of fish sera with skim milk. However, the specific immuno-reactivity of fish IgM was not reduced by such treatment. Thus, we conclude that treatment of fish sera with skim milk is useful in reducing the high background OD often observed in fish IgM detection ELISA. |
| Rights: | © 2007 Inter-Research |
| Type: | article |
| URI: | http://hdl.handle.net/2115/50879 |
| Appears in Collections: | 水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
|
Submitter: 吉水 守
|
