I have been trying to quantify total protein in a sample containing haemin using a Bradford-based assay. Results appear to be overestimated as haemin is not compatible with these type of assays. Can anyone suggest a suitable assay or a way of removing haemin from the sample? Thank you!
I must sadly and repeatedly say that the direct-use of photometer is not quantitative due to violence against Lambert-Beer's law. HPLC-photometric method gives three-fold higher values than direct-use of photometer (please see Fig.10 of file; JCB Fucoidan transport).
Furthermore, direct-photometric protein assays are interfered by many molecules, and Haemin may interfere the Bradford-based assay.
Therefore, we have developed the reliable HPLC-Surfactant-SEC protein assay method (please see file; HPLC-Surf-SEC protein determination method).
Another quantitative protein assay method is developed recently by us, which is named as PDMD (Protein-Direct-Microsequencing-Deciphering) method (please see file; HepG2 Fucoidan).
Process for the preparation of heme/hemin.
European Patent Application EP0427690
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