荧光素酶
荧光素
萤火虫协议
光发射
生物发光
化学
印版阅读器
蓝光
光化学
萤光素酶类
生物物理学
生物化学
材料科学
生物
光学
光电子学
物理
荧光
基因
动物
转染
作者
Marlene DeLuca,W. D. McElroy
出处
期刊:Methods in Enzymology
日期:1978-01-01
卷期号:: 3-15
被引量:299
标识
DOI:10.1016/0076-6879(78)57003-1
摘要
This chapter discusses purification and properties of firefly luciferase. Firefly luciferase in the presence of luciferin (d-LH2), adenosine triphosphate (ATP)-Mg, and molecular oxygen catalyzes the production of light. The color of the emitted light shows a peak at around 560 nm. This emission can be affected by temperature, pH, and metal ions. At low pH or in the presence of lead, mercury, or other heavy metals, the emission peak is shifted to red, showing an emission around 615 nm. Thus, when this system is used for the assay of ATP, it is critical to run internal controls to make sure that the light color has not been shifted. Because of the greater sensitivity of phototubes in blue in contrast to the red, a shift to red will give an apparent lower level of ATP. A dark-adapted eye can readily discern the difference in color between the control and one emitting at longer wavelengths. In crude firefly extracts and some luciferin preparations, dehydroluciferin is usually present. It is a potent inhibitor of the light reaction. The chapter describes the method of collecting and drying fireflies in order to prepare an appropriate acetone powder, the preparations and properties of a crude and partially purified luciferase and finally the preparation of crystalline firefly luciferase.
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