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.
Univ. of Wisconsin-Madison
lzhuo at facstaff.wisc.edu