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Help on reducing auto-fluorescence

Firstname Lastname Username at Waisman.Wisc.Edu
Fri Jul 30 17:49:41 EST 1999


I am seeking suggestions on reducing auto-fluorescence originated from the 
total mouse brain protein extracts, when assayed with a fluorometer (TD700, 
Turner Design) using excitation 488 nm and collection at 510-700 nm. My target 
protein for assay is the GFP expressed in transgenic brain tissues. So far I 
am able to detect significant difference in fluorescent intensity (FI) between 
the transgenic and the nontransgenic control. However the nontransgenic 
samples gave considerable autofluorescence. I would like to know if there are 
methods to reduce the auto FI to the minimum by treating/purifying total 
protein extracts proior to assay (I realize that using a narrow bandpass 
filter, e.g. 520 nm filter may also help).

Thank you for your attention.


Lang Zhuo, Ph.D.
Assistant Scientist
Waisman Center
Univ. of Wisconsin-Madison

lzhuo at facstaff.wisc.edu




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