Pavel S Gromov, University of Aarhus, Aarhus, Denmark
Julio E Celis, University of Aarhus, Aarhus, Denmark
Published online: April 2001
DOI: 10.1038/npg.els.0002689
The protein blot overlay assay is a powerful technique for identifying proteins transferred to an immobilizing matrix. It is based on the specific binding of labelled ligands with proteins of interest blotted onto the membrane, followed by detection of the complexes formed.
Keywords: protein–ligand interactions; blot overlay assay
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Figure 1. The protein blot overlay assay for a 2D protein gel.
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| References | |
| book Dejgaard K and Celis JE (1998) "Two-dimensional Northwestern blotting". In: Celis JE, Carter N, Hunter T, Shotton D, Simons K and Small JV (eds) Cell Biology. A Laboratory Handbook, vol. 4, pp. 482–486. San Diego, CA: Academic Press. | |
| Feizi T (1989) Glycoprotein oligosaccharides as recognition structures. Ciba Foundation Symposium 145: 62–74. | |
| book Gromov P and Celis JE (1998) "Blot overlay assay for the identification of GTP-binding proteins". In: Celis JE, Carter N, Hunter T, Shotton D, Simons K and Small JV (eds) Cell Biology. A Laboratory Handbook, vol. 4, pp. 454–457. San Diego, CA: Academic Press. | |
| Hausken ZE, Coghlan VM, Shafer Hastings CA, Reimann EM and Scott JD (1994) Type II regulatory subunit (RII) of the cAMP-dependent protein kinase interaction with A-kinase anchor requires isoleucine 3 and 5. Journal of Biological Chemistry 269: 24245–24251. | |
| book Hausken ZE, Coghlan VM and Scott JD (1998) "Overlay, ligand blotting and band-shift techniques to study kinase anchoring". In: Clegg RA (ed.) Protein Targeting Protocols. Methods in Molecular Biology, vol. 88, pp. 47–64. Totowa, NJ: Humana Press. | |
| book Hoffmann HJ, Gromov P and Celis JE (1998) "Calcium overlay assay". In: Celis JE, Carter N, Hunter T, Shotton D, Simons K and Small JV (eds) Cell Biology. A Laboratory Handbook, vol. 4, pp. 450–453. San Diego, CA: Academic Press. | |
| Ramos-Castaneda J, Imbert JL, Barron BL and Ramos C (1997) A 65-kDa trypsin-sensible membrane cell protein as a possible receptor for Dengue virus in cultured neuroblastoma cells. Journal of Neurovirology 6: 435–440. | |
| Further Reading | |
| book Clegg RA (ed.) (1998) Protein Targeting Protocols. Methods in Molecular Biology, vol. 88. Totowa, NJ: Humana Press. | |
| book Celis JE, Carter N, Hunter T, Shotton D, Simons K and Small JV (eds) (1998) Cell Biology. A Laboratory Handbook, vol. 4. San Diego, CA: Academic Press. | |
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