Page 140 - 006

134

+/

3. ! " ATChI 0.075 M 20 µl " 5(*
"
acetylthiocholine iodide 21.67 mg 71
/ 10 ml
4. 3! ! ' 7 (( " vortex mixer

5. (& ((
6 ! ( " & spectrophotometer ## visible '&
6/
/
)

" & 6/ 412 nm # '+
& ()(
6 ! '0
30 ' - " 3 ' & 8 /" &

1

,

' (#
' 5 5( (".
*
!
Ellman et
al. (1961) ( 7

Enzyme activity = 574 x ∆A mUnit/ml
4
= 2.87 x 10 x ∆A mUnit/g (tissue)
4
= 2.87 x 10 x ∆A x 100/P mUnit/g (protein)

6/ ∆A : & absorbance ' / /" 5 '
P : , Y , 67 "6/

R
2) # $M $IO # $ 8 [x esterase
& ,

7
+/
( 7 (Chotimanothum 2000)
1. 1 67 "6/ ! " buffer phosphate (pH 7.5, 0.1
M˜

67
6/
M) ! " 50 mg buffer 1 ml polyvinul polypyrrolidone (PVPP)
& +/ +/ 71
67 "6/ #( "( ' / "Y

/
6/
2. ! "'5( ! ( microtube 5 /" ( " &
1
1
centrifuge ( "& Y 18,000 # ' (10,000 g) 0 -) 4°C 5 '
/

/
)

3. ((! "'5( 50 µl ! (' potassium phosphate buffer

(pH 7.5, 0.1 M) 2.9 ml 50 µl PNPA (paranitrophenyl acetate) (" "# '"#
#!

6
J & 50 µl potassium phosphate buffer (' potassium phosphate
/
buffer 2.9 ml EDTA (ethylene diamine tetraacetic acid), GSH (glutathione, pH 7.5)
50 µl PNPA
4. 3! ! ( ( "
6/

/
5. * (& ()(
6 ! ("' ' ( " & spectrophotometer '&

" & 6/ 400 nm '
30 ' 3 ' & 8 /" &
'
0

("! ! 6 paranitrophenol *

hydrolysis ! paranitrophenyl acetate

,
1
,

5 ! ! ! H (#
' 5 ! !5( Mackness et
al. 1983
3) # $M $IO # $ 8 [x glutathione S  transferase
R
+/
7
& ,( 7 (Chotimanothum 2000)