Back to Search
Start Over
Reusable on-plate immunoprecipitation method with covalently immobilized antibodies on a protein G covered microtiter plate.
- Source :
-
Journal of Immunological Methods . Aug2020, Vol. 483, pN.PAG-N.PAG. 1p. - Publication Year :
- 2020
-
Abstract
- Covalent immobilization of antibodies to protein G beads is a basic molecular biology method, although the beads present poor recovery results. Our aim was to reuse the immobilized antibody-protein G complex on a very small scale, therefore we optimized the crosslinking procedure to be used on the wells of a standard 96-well microplate. The method used involves the affinity binding of the antibody to the protein G surface, followed by the immobilization step using different crosslinking reagents, DMP and BS3, quenching the crosslinking reaction, and binding the antibody-specific antigen. By scaling down the procedure, we were able to reuse the anti-EGFR crosslinked wells more than 20 times. This method can be used to perform assays on a wide range of solid supports containing the protein G in an immobilized form, including functionalized nanosensors, for immunoprecipitation, protein and cell lysate purification, target protein enrichment. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00221759
- Volume :
- 483
- Database :
- Academic Search Index
- Journal :
- Journal of Immunological Methods
- Publication Type :
- Academic Journal
- Accession number :
- 144460059
- Full Text :
- https://doi.org/10.1016/j.jim.2020.112812