protein absorbance wavelength? - (Sep/15/2008 )
hello,
im doing a protein quantification assay (using biorad reagents with BSA as my protein standard). i was wondering what absorbance should i take my readings at. the protocol from biorad says 750 (650-750 is ok). other places online ive seen 595 and one time i saw 405. anyone wanna clarify? thanks!!
-medstudent1-
QUOTE (medstudent1 @ Sep 16 2008, 10:02 AM)
hello,
im doing a protein quantification assay (using biorad reagents with BSA as my protein standard). i was wondering what absorbance should i take my readings at. the protocol from biorad says 750 (650-750 is ok). other places online ive seen 595 and one time i saw 405. anyone wanna clarify? thanks!!
im doing a protein quantification assay (using biorad reagents with BSA as my protein standard). i was wondering what absorbance should i take my readings at. the protocol from biorad says 750 (650-750 is ok). other places online ive seen 595 and one time i saw 405. anyone wanna clarify? thanks!!
Various assays use different chemistries to detect and quantify the protein content, so you need different wavelengths to detect the final products. Lowry detects at 750 nm, Bradford uses 595, while the BCA assay (a derivative of the Lowry) uses 562 nm.
-swanny-