1、 -Cu2+ 540nm 0.5g/L10g/L TrisGood10% 1m?NaOH Lowry Cu+ - 20mg/L 400mg/L pH- pH10Cu2+- - Cu2+- 280nm 280nm0.10.5mg /ml260nm 280nm1 260nm1 0.1%280nm0.52.5 230nm0.1%2 25nm215nm5.011.7280nm0.58230nm 215nm225 nm10l/ml100l/ml pH ABradford ()- (Biuret)Folin?(Lowry) 1976Bradford(Bradford) - G- 250(lma x)465
2、nm595nm- () 595nmA595 Bradford: (1)Lowry1mg - Low ry (2)5 2 1520 Lowry (3)LowryK+Na+Mg2+Tris EDTA : (1)Bradfo rd g? (2):Triton X- 100(SDS)0.1NNaOH(0.1NLowary ) (3)Beer () 1. : (1) g?(BSA)1.0mg/ml0.1mg/ml (2)G-250:100mgG-25050ml 95%120ml 85%1 2. : (1) (2) (3)16 () 1. (1)1613 1.0mg/ml :00.010.020.040.
3、060.080.1ml 0.1ml5.0mlG- 250( )8910 (2)25 595nmA59510.1mlH2O5.0m lG-250 :() 95% (3)(mg)A595 A595 0.5mg/mlA5950.50 2. (10- 100mg/ml)()0.5ml1.0ml 0.5ml1.0ml H2OG- 2505.0ml595nm0. 05mg/mlA595 . Boug uer1729Lambert1760 : T=I/I0=e-kb I0Iekb (cm) Bouguer Beer-Bouguer: T=I/I0=e-kbc c(g/Lmg/L)10 : A=-logT=-
4、log(I/I0)=log(I0/I)=bc A = BeerA=kbccg/L bcmKa/ L/(gcm,A=kbcA=abcA=kbc1 mol/L,b1cmk/ L/(molcmA=kbcA=c1mol /L NADH260nm15000 260NADH=15103340nm6220340NADH=6.22103 A=kbccw/vbcmkE% A=kbcA=E%bc aE B . 280nm 238nm NH42SO4 - pH pHpHpH 1. 280nm 280nm 280nm 280nmA28 00.11.0mg/ml1cm 1mg/mlA2801.0 A11cm A11cm
5、1%1cm A11cm, = (A28010 )/ A11cm,280nm (mg/ml) (Q 1%?10mg/ml) 2. 280nm260nm 280nm10 260nm260nm260nm 280nm2280nm260nm A280/A260 ? 1.8 A280/A260 ? 0.5 A280A260 - =1.45A280-0.74A260 (mg/ml) - 3. 215nm225nm 280nm215nm 225nm 20100 mg/ml5.0ml215nm225nm D= A215 -A225 D 20100mg/mlN aClNH42SO40.1MTris 0.1N NaOH, 0.1M215nm 0.005M 4. 238nm 50500mg/ml5.0ml2 38nmA238A238, 238nm 280nm
