Table 3.
Summary details of the protocols for the here-biomarker determinations.
| Biomarkers | Sample | Tissue preparation |
Biomarker determinations |
|||
|---|---|---|---|---|---|---|
| Homogenization Centrifugation |
Dilution factor | Spectrophotometric readings (nm) | Result expression units | References | ||
| PROT |
All |
same to the corresponding biomarker |
1:20 |
595 |
mg protein/body weight |
[53] |
| SOD | Body (2 fish per replicate) |
2.5 mL - phosphate buffer (50 mM, pH 7.0) with Triton X-100 (0.1%) 15,000 rpm; 10 min; 4 °C |
1:15 | 500 | units min/mg/protein | [54] |
| CAT | 1:100 | 240 | μmol/min/mg protein | [55] | ||
| GPx | 1:2 | 340 | mmol/min /mg protein |
[56] | ||
| GRed | 1:2 | 340 | μmol/min/mg protein | [57] | ||
| GSH | 1:20 | 412 | μg/mg protein | [58] | ||
| GSTs | 1:20 | 340 | mmol/min /mg protein |
[59] | ||
| TBARS |
1:10 |
535 |
mmol/mg protein |
[60] |
||
| GLY |
Body (1 fish per replicate) |
2.5 mL - phosphate buffer (50 mM, pH 7.0) with Triton X-100 (0.1%) 4000 rpm; 10 min; 4 °C |
1:1 |
490 |
μg/mg protein |
[61] |
| LIP |
Body (1 fish per replicate) |
4 mL - chloroform: methanol (2:1) mixture 3000 rpm; 10 min |
1:1 |
* |
mmol/mg protein |
[62] |
| LDH |
Body (1 fish per replicate) |
2.5 mL - TRIS buffer (0.1 M, pH = 7.2) 6000 rpm; 3 min; 4 °C |
1:15 |
340 |
mmol/min /mg protein |
[63] |
| AChE | Head (1 fish per replicate) | 1.5 mL - phosphate buffer (0.1 M, pH 7.2) 6000 rpm; 5 min; 4 °C |
1:20 | 412 | nmol/min/mg protein | [64] |
*Extraction procedure through the biphasic solvent system consisting of chloroform/methanol/water. The results (% of lipids) were obtained by the difference between the weight of tubes before and after the lipids extraction