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V6=0@>4=89 V=45:A GCB;82>ABV (') 2:070=> 2 0A?>@BV 4> =01>@C. 2. >=B@>;L=0 ?;07<0 (;V>DV;L=> 28ACH5=0 :>=B@>;L=0 ?;07<0 :@>2V ;N48=8), =0 1 ml (<;) - 1 D;. 3. =AB@C:FVO 7 28:>@8AB0==O. 4. 0A?>@B =0;VB8G=V E0@0:B5@8AB8:8 >5DVFVT=B 20@V0FVW @57C;LB0BV2 287=0G5==O ?@>B@><1V=>2>3> G0AC =5 ?>28=5= ?5@528IC20B8 10%. >?CAB8<89 @>7:84 @57C;LB0BV2 287=0G5==O ?@>B@><1V=>2>3> G0AC 2 >4=V9 ?@>1V ?;07<8 :@>2V @V7=8<8 =01>@0<8 >4=VTW A5@VW =5 ?>28=5= ?5@528IC20B8 10%1238556 01V@ 0;0=V=-0<V=>B@0=AD5@078;0=V=0<V=>B@0=AD5@070 (ALT/GPT) 1 x 200 <;HB3 5035=B8 ?>28==V 1CB8 ?@87=0G5=V 4;O 28<V@N20==O :>=F5=B@0FVW " 2 A8@>20BFV 01> ?;07<V :@>2V ;N48=8, H;OE>< 287=0G5==O 0:B82=>ABV ALT 70 H284:VABN 7<5=H5==O NADH. 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(?5@5=5A5==O 0<V=>3@C?8 2V4 0A?0@B0BC 4> 2->:A>3;NB0@0BC, CB2>@NNG8 >:A0;0F5B0B V 3;NB0<0B.) !:;04: . 5035=B 1 E 160 <; : "@VA 121 <<>;L/;, L-0A?0@B0B 362 <<>;L/;, <0;0B453V4@>35=070 >460 4/;, ;0:B0B453V4@>35=070 > 660 4/;, pH 7.8. . 5035=B 1 E 40 <; : NADH 1.9 <<>;L/;, 2->:A83;NB0@0B 75 <<>;L/;, 3V4@>E;>@84 =0B@VN 148 <<>;L/;, 0784 =0B@VN 9.5 3/;. 560 287=0G5==O =5 28I5 1.67 4/;. 560 ;V=V9=>ABV =5 <5=H5 800 4/;.1433165 01>@8 ;C6=>W D>AD0B078C6=0 D>AD0B070 (ALP)-AMP 1 x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x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x 100 <; 50:B82. VB@8B =0B@VN 11.6 <<>;L/;.  # ( /) AD. 400 <; 5:0B82. !C;LD0=V;>20 :8A;>B0 35 <<>;L/;, A>;O=0 :8A;>B0 0.24 <>;L/;. BD. 1 x 100 <; 50:B82. VB@8B =0B@VN 3.5 <<>;L/;. 560 287=0G5==O 4;O 7030;L=>3> 1V;V@C1V=C =5 28I5 0.03 <3/4; = 0.51 <:<>;L/;. 560 ;V=V9=>ABV 4;O 7030;L=>3> 1V;V@C1V=C =5 <5=H5 20 <3/4; = 343 <:<>;L/;. 560 287=0G5==O 4;O ?@O<>3> 1V;V@C1V=C =5 28I5 0.02 <3/4; = 0.34 <:<>;L/;. V=V9=VABL 4;O ?@O<>3> 1V;V@C1V=C =5 <5=H5 20 <3/4; = 343 <:<>;L/;.1741823 030B>:><?>=5=B=89 :>=B@>;L :;V=VG=>W EV<VW, 0=0;V7>20=89, =>@<0;L=89V>EV<VG=0 :>=B@>;L=0 A8@>20B:0 (HUMAN) 5%5 <;HB1@87=0G5=0 4;O :>=B@>;N O:>ABV B0 ?>28==0 <0B8 @5:><5=4>20=V 4V0?07>=8 4>?CAB8<8E 7=0G5=L. 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HEPES 50 <<>;L/;, E;>@84 :0;LFVN 0.075 <<>;L/;, E;>@84 <03=VN 13 <<>;L/;, a 3;N:>784070 > 4 4/<;, pH 7.1 B. 5035=B : 1 x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ml (<;) , - 1 D;. 2. =AB@C:FVO 7 28:>@8AB0==O. 3. 0A?>@B. =0;VB8G=V E0@0:B5@8AB8:8 ?>28==V 1CB8: >5DVFVT=B 20@V0FVW @57C;LB0BV2 287=0G5==O 0B5AB>20=8E ?>:07=8:V2 2 :0;V1@C20;L=V9 ?;07<V =5 ?5@528IC20B8 5%. >?CAB8<5 2V4E8;5==O ?>:07=8:V2 2 :0;V1@C20;L=V9 ?;07<V 2V4 0B5AB>20=>3> 7=0G5==O =5 ?5@528IC20B8 5%. >?CAB8<89 @>7:84 @57C;LB0BV2 287=0G5==O 0B5AB>20=8E ?>:07=8:V2 2 @V7=8E @5035=B0E >4=VTW A5@VW =5 ?5@528IC20B8 5%.2130499 01V@ @5035=BV2 4;O 28<V@N0==O !-@50:B82=>3> 1V;:0! -;0B5:A-B5ABHB1001V@ 70AB>A>2CTBLAO 4;O 28O2;5==O !-@50:B82=>3> 1V;:C 2 A8@>20BFV :@>2V ;N48=8 ! -;0B5:A-B5AB. 01V@ ?>28=5= 1CB8 @>7@0E>20=89 =0 200 287=0G5=L 7 C@0EC20==O< E>;>AB8E ?@>1 ?@8 28B@0BV @>1>G>3> @>7G8=C 2V4?>2V4=> FVTW <5B>48:8. 01V@ ?>28=5= A:;040B8AO: 1. 5035=B 1. 0B5:A=0 ACA?5=7VO, 2 <; (1 HB.) 2. 5035=B 2. >7G8==8:, 14 <; (1 HB.) 3. 5035=B 3. >78B82=89 :>=B@>;L, O:89 <VAB8BL !  1V;LH 6 <3/;, 0.2 <; (1 HB.) 4. 5035=B 4. 530B82=89 :>=B@>;L, O:89 <VAB8BL !  <5=H 6 <3/;, 0.2 <; (1 HB.) 5. 0;8G:8 4;O @>7<VHC20==O A8@>20B>: (100 HB.) 6. 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