For the Determination of SGOT (AST) Activity in Serum

1. • ABDUL BATEN

2.  Test tube
 Test tube stand
 Micropipette
 Micro Tips
 Blood sample
 SGOT kit
 Syringe
 Vacutainer tube
 Tourniquet

3.  SGOT (AST) catalyzes the transfer of amino
group between L-Asparate and a-Ketoglutarate
to form oxaloacetate and Glutamate.
 The oxaloacetate formed reacts with NADH in
the presence of Malate Dehydrogenase to form
NAD.
 The rate of oxidation of NADH to NAD is
measured as a decrease in absorbance which is
proportional to the SGOT(AST) activity in the
sample.

4. L- Asparate + α- ketoglutarate L- Glutamate + oxaloacetate
Oxaloacetate + NADH +H+ L- malate + NAD+

5.  WORKING REAGENT:- mix 4 parts of R1 + 1 part
of R2
 Pipette into clean dry test tubes labeled as
Standard (S), and Test (T): T
 Working Reagent:- 1000μl (1ml)
 Specimen :- 100μl (0.10m)
# Mix well and read the initial absorbance A after 1
min. and repeat 0 the absorbance reading after
1, 2 & 3 minutes at 37oC

6.  A/min X factor= AST activity( U/L)
 Factor= 1746
 NORMAL VALUES: (Serum)
 Males : ≤37 U/L
 Females: ≤ 31 U/L
 CLINICAL SIGNIFICANCE:
INCREASES: Increased levels are found in hepatobiliary disease.