Abstract
The use of Gaussia luciferase in a multiplex assay can have several advantages over the singleplex method for an experimental setup. Issues such as intersample variability, screening purposes, efficiency, and in vivo applications can be addressed using a multiplex assay. Here we describe a functional reporter multiplex method using Gaussia luciferase fused to epitope tags to identify the different reporters that are expressed. Tag specific antibodies are used to bind and separate the tagged luciferase reporters.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Badr CE, Tannous BA (2011) Bioluminescence imaging: progress and applications. Trends Biotechnol 3:1–10. doi:10.1016/j.tibtech. 2011.06.010
Stratowa C, Audette M (1995) Transcriptional regulation of the human intercellular adhesion molecule-1 gene: a short overview. Immunobiology 193:293–304. doi:10.1016/S0171-2985(11)80558-9
Wrana JL, Attisano L, Wieser R et al (1994) Mechanism of activation of the TGF-beta receptor. Nature 370:341–347. doi:10.1038/370341a0
Subramaniam D, Natarajan G, Ramalingam S et al (2008) Translation inhibition during cell cycle arrest and apoptosis: Mcl-1 is a novel target for RNA binding protein CUGBP2. Am J Physiol Gastrointest Liver Physiol 294:G1025–G1032. doi:10.1152/ajpgi.00602.2007
Niers JM, Kerami M, Pike L et al (2011) Multimodal in vivo imaging and blood monitoring of intrinsic and extrinsic apoptosis. Mol Ther 19:1090–1096
De Wet JR, Wood KV, DeLuca M et al (1987) Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol 7:725–737
Loening AM, Wu AM, Gambhir SS (2007) Red-shifted Renilla reniformis luciferase variants for imaging in living subjects. Nat Methods 4:641–643. doi:10.1038/nmeth1070
Tannous BA, Kim D-E, Fernandez JL et al (2005) Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo. Mol Ther 11:435–443. doi:10.1016/j.ymthe.2004.10.016
McNabb DS, Reed R, Marciniak RA (2005) Dual luciferase assay system for rapid assessment of gene expression in Saccharomyces cerevisiae. Eukaryot Cell 4:1539–1549. doi:10.1128/EC.4.9.1539-1549.2005
Van Rijn S, Nilsson J, Noske DP et al (2013) Functional multiplex reporter assay using tagged Gaussia luciferase. Sci Rep 3:1046. doi:10.1038/srep01046
Ford CF, Suominen I, Glatz CE (1991) Fusion tails for the recovery and purification of recombinant proteins. Protein Expr Purif 2:95–107
Dull T, Zufferey R, Kelly M et al (1998) A third-generation lentivirus vector with a conditional packaging system. J Virol 72: 8463–8471
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
van Rijn, S., Würdinger, T., Nilsson, J. (2014). Multiplex Functional Bioluminescent Reporters Using Gaussia Luciferase Fused to Epitope Tags in an Immunobinding Assay. In: Badr, C. (eds) Bioluminescent Imaging. Methods in Molecular Biology, vol 1098. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-718-1_18
Download citation
DOI: https://doi.org/10.1007/978-1-62703-718-1_18
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-717-4
Online ISBN: 978-1-62703-718-1
eBook Packages: Springer Protocols