The ALT and AST activity in serum of rats were determined using commercial kits produced by BioMar (Gliwice, Poland). Normal and pathological serum were used for quality control. Cytochrome P450 1A (CYP1A) subfamily in mammals consists of 2 isoforms, CYP1A1 and CYP1A2. Activity of the enzymes in liver of rats was measured using the kinetic method described by Dey et al. [26 (link)] and Zamaratskaia et al. [27 (link)] with ethoxyresorufin (CAS #5725-91-7, Sigma-Aldrich #46121-5MG-F, Saint Louis, USA) and methoxyresorufin (CAS #5725-89-3, Sigma-Aldrich #69125-5MG, Saint Louis, USA) as reagents. The fluorescence of both enzymatic reactions was measured at 535 nm and 590 nm as excitation/emission wave lengths, protein concentration was analyzed with Bradford reagent. Bovine serum albumin was used as standard.
Protocol detail
Serum Enzyme Activity Assay in Rats
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Bovine serum albumin
Cyp1a1
Cyp1a2
Cytochrome p450
Enzymatic
Enzymes activity
Ethoxyresorufin
Fluorescence
Isoforms
Kinetic
Liver
Mammals
Protein
Rats
Serum
Corresponding Organization :
Other organizations : Nofer Institute of Occupational Medicine
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Variable analysis
independent variables
- None explicitly mentioned
- ALT (alanine aminotransferase) activity in serum of rats
- AST (aspartate aminotransferase) activity in serum of rats
- Activity of cytochrome P450 1A (CYP1A) subfamily enzymes in the liver of rats
- Normal serum used for quality control
- Pathological serum used for quality control
- Protein concentration analyzed with Bradford reagent
- Bovine serum albumin used as standard
- Positive control: Normal serum
- Negative control: Pathological serum
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