nh lng protein theo phng php Bradford (1976) Nguyn tc Cc protein khi phn ng vi coomassie (coomassie brilliant blue-CBB)

) s hnh thnh hp cht mu c kh nng hp th nh sng bc sng 595 nm, cng mu t l vi nng protein trong dung dch. Phng php ny c nhy cao cho php pht hin t 5 - 200g protein, d thc hin v tit kim thi gian. Ha cht Thuc nhum Bradford: ha tan 50 mg thuc nhum coomassie brilliant blue G250 vo 23,5 g ethanol 96o, thm 42,5 g H3PO4 85% v chnh ti 500 ml bng nc ct. Dung dch albumin 0.1 mg/ml: cn 10 mg albumin pha vi nc ct thnh 100 ml. Dch mu protein: 1 ml cho mt phn ng Tin hnh th nghim Xy dng ng chun albumin Ht 1ml dung dch protein chun cc nng t 10-80g, thm vo 2 ml thuc nhum Bradford, lc u. Sau em ly tm 10 pht em o OD ti bc sng 595 nm. V th biu din s bin thin mt quang (OD) theo nng protein chun (g/ml) . Xc nh hm lng protein trong mu Tng t ht 1ml protein cn phn tch, thm vo 2 ml thuc nhum Bradford, yn 10 pht, em o OD ti bc sng 595nm. T ng chun suy ra hm lng protein cn phn tch (OD595 * pha long ca mu)