Active Wnt signaling results in other effects on cells such as phosphorylation of Dishevelled , or phosphorylation of the LRP tail Tamai To activate Wnt signaling, one can add Wnt protein , either in a purified form or as conditioned medium to cells.
Active Wnt protein can be obtained from several companies. An alternative to Wnt protein is the use of Wnt Surrogates , soluble proteins that bind to and activate Wnt receptors Janda et al, , T ao et al. Comparing the Wnt conditioned medium to medium from control L cells is a reasonable way of controling for specificity but it should be realized that Wnt expression can lead to the induction of genes encoding other secreted factors such as FGFs, see target genes and that the conditioned medium may therefore contain such factors.
Whether cells respond to the particular Wnt will depend on the receptors expressed. There is something known about Wnt-Frizzled receptor specificity. How cells will respond to Wnt signaling depends very much on the state of the cell; most of the Wnt target gene s are cell type specific and only activated in certain tissues. Several other inhibitors of GSK3 have been described Meijer To measure luciferase activity of the cell lysates, you will need a multiwell plate or a tube containing cell lysates and a luminometer.
This apparatus detects the photon emission produced from the luciferase reaction and the unit of measurement comes out as relative light units RLU. The injector dispenses a substrate into the cell lysate and the reading of the wells starts immediately to minimize the possibility of diminishing signal Smale, A Crash Course on Luciferase Assays. Badr, C.
Bioluminescence imaging: progress and applications. Trends in Biotechnology, 29 12 , — Brasier, A. ScienceDirect; Academic Press. Carter, M. Shieh eds. Cheng, Z. Current Chemical Genomics, 4, 84— Gong, D. Journal of Biological Chemistry, 8 , — Hampf, M. A protocol for combined Photinus and Renilla luciferase quantification compatible with protein assays.
Analytical Biochemistry, 1 , 94— He, M. Matta, H. Development and characterization of a novel luciferase based cytotoxicity assay. Scientific Reports, 8. Min, J. Journal of Clinical Medicine, 9 8 , The best answers are voted up and rise to the top. Stack Overflow for Teams — Collaborate and share knowledge with a private group.
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