Is anyone here using the Thermo Fisher Nano Orange kit for protein quantification, and detecting the fluorescent signal on the Promega Quantus?
We are struggling to make sense of the standard curve data we're generating with the BSA supplied. The product insert for the Nano Orange says it is linear in the 10 ng/mL to 10 ug/mL range, but our Quantus is definitely not giving linear readings below 10 ug/mL. There is almost no difference between the blank, the 0.1 ug/mL and the 1 ug/mL BSA using the raw RFU. Seems our Nano Orange is not becoming bright enough for our Quantus to detect, or maybe the Quantus simply is not sensitive enough to read, even at the top end of the Nano Orange scale.
While we wait to hear back from the various suppliers, I thought I would ask you all ... what level of RFU should we be seeing for 10 ug/mL of BSA?
Thanks in advance!
Melanie
Melanie J Mcconnell
Quantus fluorometer Promega, USA is designed to measure the concentration of nucleic acids (DNA and RNA), not proteins.
Thanks, Yuri - Promega don't sell a kit for protein but you can set up your own custom programs, as it has 2 fluorometer channels - 'blue' and 'red' with specific excitation and emission. The Nano Orange has the right parameters to work on the Quantus, and Promega have a protocol for it.
Melanie J Mcconnell
I wish you success. DNA and RNA have many heterocycles, but protein has few.
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