ReadiUse™ NADP Regenerating Kit
Example protocol
AT A GLANCE
Important notes
Thaw all the components at room temperature before use.
PREPARATION OF STOCK SOLUTION
1. NADPH solution (40X):
Add 0.5 mL H2O into one vial of NADPH (Component D) to make 40X NADPH solution. Note: One vial of NADPH is for making 10 mL NADP Regenerating Solution.
2. NADP Regenerating Enzyme Mix solution (400X):
Add 100 µL H2O into the vial of NADP Regenerating Enzyme Mix (Component C) to make 400X NADP Regenerating Enzyme Mix solution. Protect from light.
PREPARATION OF WORKING SOLUTION
Add 5 mL of NADP Regenerating Buffer I (Component A), 5 mL of NADP Regenerating Buffer II (Component B), 0.5 mL of 40X NADPH solution, and 50 μL of 400X NADP Regenerating Enzyme Mix solution to make a total volume of 10.55 mL 2X NADP regenerating solution. Keep from light. Note: Activated NADP regenerating solution is not stable, and should be prepared freshly before use. 10 mL of NADP Regenerating Solution is enough for 1 plate.
SAMPLE EXPERIMENTAL PROTOCOL
- Add equal volume of 2X NADP Regenerating solution into the desired assay system.
Citations
Authors: Song, Minmin and Gorzalski, Andrew and Nguyen, Trang T and Liu, Xibei and Jeffrey, Christopher and Blomquist, Gary J and Tittiger, Claus
Journal: Journal of chemical ecology (2014): 181--189
Authors: Song, Minmin and Delaplain, Patrick and Nguyen, Trang T and Liu, Xibei and Wickenberg, Leah and Jeffrey, Christopher and Blomquist, Gary J and Tittiger, Claus
Journal: Insect biochemistry and molecular biology (2014): 73--80
Authors: Song, Minmin and Kim, Amy C and Gorzalski, Andrew J and MacLean, Marina and Young, Sharon and Ginzel, Matthew D and Blomquist, Gary J and Tittiger, Claus
Journal: Insect biochemistry and molecular biology (2013): 336--343
Authors: Qiu, Yue and Tittiger, Claus and Wicker-Thomas, Claude and Le Goff, Gaelle and Young, Sharon and Wajnberg, Eric and Fricaux, Thierry and Taquet, Nathalie and Blomquist, Gary J and Feyereisen, René
Journal: Proceedings of the National Academy of Sciences (2012): 14858--14863