Elisa Michelini, Luca Cevenini, Laura Mezzanotte, Aldo Roda's Bioluminescence: Methods and Protocols PDF
By Elisa Michelini, Luca Cevenini, Laura Mezzanotte, Aldo Roda (auth.), Preston B. Rich, Christelle Douillet (eds.)
Through the learn and alertness of bioluminescence, scientists have painstakingly harnessed a strong device that permits us to hunt a deeper realizing of the complicated mechanisms underpinning such a lot of very important biologic platforms. during this absolutely revised and up-to-date moment variation of Bioluminescence: tools and Protocols, specialist researchers give a contribution a readable and utilitarian compilation of the most recent and so much cutting edge concepts that experience emerged during this swiftly increasing and steadily various box together with easy methods to investigate telephone trafficking, protein-protein interactions, intracellular signaling, and apoptosis. additionally commencing up the prospect to imagine and quantify organic mechanisms in real-time and in in vivo settings, the amount additionally describes the in vivo learn of bacterial or viral infections, transplanted cells, stem cells proliferation, vascular circulation, and tumors. Written within the hugely winning Methods in Molecular Biology™ sequence structure, chapters contain short introductions to their respective subject matters, lists of the required fabrics, gear, and reagents, step by step, easily reproducible laboratory protocols, and notes on troubleshooting and warding off identified pitfalls.
Authoritative and state-of-the-art, Bioluminescence: tools and Protocols, moment version provides protocols which are designated sufficient to be and tailored through clinical groups who've no prior services in bioluminescence as a way to foster the doubtless a number of breakthroughs and new functions from easy to utilized technology and drugs that needs to remain developed.
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Additional resources for Bioluminescence: Methods and Protocols
Other doses of luciferin may be used. Caliper recommends a dose of 150 mg/kg for mice with its lumina imaging system. Our experience favors the higher dose. Caliper Lifesciences recommends 10–15 min between injection and imaging. 7. The exposure time may be altered to avoid overexposing intense signals or to detect weak signals. In practice, we may use anywhere from 1 to 30 min. Many investigators like to maintain a constant imaging time, where 5 min is typical. 8. The macros are really outside the scope of this chapter and interested readers are referred to (8).
Quantification of ATP using luciferin/luciferase. (A) Anions greatly interfere with the luciferase reaction. Calibration curves of ATP were performed by adding 30 ml of the indicated ATP concentrations to 300 ml H2O or 154 mM NaCl. Values are the mean – SEM of two separate experiments, n = 3. (B) Serum components decrease ATP detection. ATP was diluted at the indicated concentrations in H2O, HBSS+, MEM, or MEM supplemented with 10% FBS. A 30-mL aliquot was collected and added to a 5-mL test tube containing 300 mL of water.
And Rice, K. G. (2006) Quantitative bioluminescence imaging of transgene expression in vivo. Anal Biochem 355, 90–94. Thompson, J. , Hayes, L. , and Lloyd, D. B. (1991) Modulation of firefly luciferase 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. stability and impact on studies of gene regulation. Gene 103, 171–177. Burgos, J. , Moats, R. , Kohn, D. , Nelson, M. , and Laug, W. E. (2003) Time course of bioluminescent signal in orthotopic and heterotopic brain tumors in nude mice. BioTechniques 34, 1184–1188.
Bioluminescence: Methods and Protocols by Elisa Michelini, Luca Cevenini, Laura Mezzanotte, Aldo Roda (auth.), Preston B. Rich, Christelle Douillet (eds.)