Blood samples harvested from euthanised female offspring rats were centrifuged at 1500× g and 4 °C for 10 min, and the resulting supernatant (plasma) was collected for subsequent experiments. In this study, the concentrations of triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were determined using the Free Fatty Acid Quantification Assay Kit (Abcam, Cambridge, UK) [91 ], the Cholesterol/Cholesteryl Ester Quantitation Assay Kit (Abcam, Cambridge, UK) [92 ], and the HDL and LDL/VLDL Cholesterol Assay Kit (Abcam, Cambridge, UK) [93 ], while the plasma concentration of leptin was measured using the Leptin Rat ELISA Kit (Thermo Fisher Scientific, Waltham, MA, USA) [94 ] according to the manufacturer’s instructions.
Leptin rat elisa kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Leptin Rat ELISA Kit is a quantitative assay designed to measure the concentration of leptin in rat serum, plasma, and other biological fluids. The kit utilizes the enzyme-linked immunosorbent assay (ELISA) technique to provide accurate and reliable results.
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4 protocols using leptin rat elisa kit
1
Serum Lipid and Leptin Analysis in Rat Offspring
2
Comprehensive Biomarker Analysis in Rats
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The plasma levels of leptin (Leptin Rat ELISA Kit; Thermo Fisher Scientific, Inc., Waltham, MA, USA), troponin-I (Rat Cardiac Troponin I ELISA Kit (ab246529); Cambridge, MA, USA), FGF-21 (RayBio® Mouse FGF-21 ELISA Kit; Peachtree Corners, GA, USA), IL-6 (RayBio® Rat IL-6 ELISA Kit; Peachtree Corners, GA, USA), and TNF-alpha (RayBio® Rat TNF-alpha ELISA Kit; Peachtree Corners, GA, USA) were measured using commercial ELISA kits, using specific ELISA kits, according to the manufacturer's instructions. Plasma glucose, total cholesterol, HDL, LDL, and triglyceride concentrations were determined using commercially available kits (Olvex Diagnosticum; Saint-Petersburg, Russia).
3
Plasma Leptin Levels in Cocaine Withdrawal
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- Baseline blood (before cocaine): Retroorbital bleeds were performed under anesthesia with the eye numbed by a topical ophthalmic anesthetic (proparacaine hydrochloride).
- Terminal (acute cocaine withdrawal: euthanized 18 h in withdrawal after a LgA session or protracted cocaine abstinence: euthanized∼3–4 weeks after the last cocaine intake during SA sessions): Blood was collected through cardiac puncture after CO2 inhalation.
Fresh blood was collected in EDTA coated tubes to avoid coagulation and was centrifuged at 2,000 g at room temperature (RT) for 10 min. The supernatant plasma was immediately transferred per 500 μl aliquots into fresh tubes, scored for quality on a scale from 0 to 6, snap-frozen on dry ice, and stored at –80°C. Frozen aliquots were thawed on ice and analyzed using a Leptin Rat ELISA kit (Invitrogen, Waltham, MA, United States).
- Terminal (acute cocaine withdrawal: euthanized 18 h in withdrawal after a LgA session or protracted cocaine abstinence: euthanized∼3–4 weeks after the last cocaine intake during SA sessions): Blood was collected through cardiac puncture after CO2 inhalation.
Fresh blood was collected in EDTA coated tubes to avoid coagulation and was centrifuged at 2,000 g at room temperature (RT) for 10 min. The supernatant plasma was immediately transferred per 500 μl aliquots into fresh tubes, scored for quality on a scale from 0 to 6, snap-frozen on dry ice, and stored at –80°C. Frozen aliquots were thawed on ice and analyzed using a Leptin Rat ELISA kit (Invitrogen, Waltham, MA, United States).
4
Cocaine-Induced Plasma Leptin Dynamics
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-Baseline blood (before ShA): Retroorbital bleeds were performed under anesthesia with the eye numbed by a topical ophthalmic anesthetic (proparacaine hydrochloride).
-Terminal: Blood was collected through cardiac puncture after CO2 inhalation. Each cohort was divided into three final endpoints. Intoxication: euthanized with drug on board after a 3h SA session. Withdrawal: euthanized 18h in withdrawal after a LgA session. Abstinence: euthanized ~3-4 weeks after the last cocaine intake during SA sessions.
Fresh blood was collected in EDTA coated tubes to avoid coagulation and was centrifuged at 2000 g at room temperature (RT) for 10 min. The supernatant plasma was immediately transferred per 500 µl aliquots into fresh tubes, scored for quality on a scale from 0-6, snap frozen on dry ice and stored at -80°C. Frozen aliquots were thawed on ice and analyzed using a Leptin Rat ELISA kit (Invitrogen).
-Terminal: Blood was collected through cardiac puncture after CO2 inhalation. Each cohort was divided into three final endpoints. Intoxication: euthanized with drug on board after a 3h SA session. Withdrawal: euthanized 18h in withdrawal after a LgA session. Abstinence: euthanized ~3-4 weeks after the last cocaine intake during SA sessions.
Fresh blood was collected in EDTA coated tubes to avoid coagulation and was centrifuged at 2000 g at room temperature (RT) for 10 min. The supernatant plasma was immediately transferred per 500 µl aliquots into fresh tubes, scored for quality on a scale from 0-6, snap frozen on dry ice and stored at -80°C. Frozen aliquots were thawed on ice and analyzed using a Leptin Rat ELISA kit (Invitrogen).
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