Brain FFA content was measured using the FFA quantification kit from BioVision following the manufacturer’s instructions. Fluorescence was measured using Corning black 96-well polypropylene assay plates and the Microplate Multimode Reader with green fluorescence module (Ex 525 nm; Em 580–640 nm). FFA concentrations were calculated using a standard curve for palmitic acid ranging from 0 to 0.02 nmol/µL.
Ffa quantification kit
Manufactured by Abcam
Sourced in United States, United Kingdom
The FFA quantification kit is a laboratory tool designed to quantify free fatty acids (FFAs) in various sample types. This kit provides a reliable and efficient method for measuring FFA levels, which is essential for research and analysis in fields such as biochemistry, nutrition, and metabolic studies.
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Lab products found in correlation
Infinity cholesterol reagent, by Thermo Fisher Scientific (1 mentions)
Infinity tg reagent, by Thermo Fisher Scientific (1 mentions)
Onetouch ultra 2 blood glucose meter, by LifeScan (1 mentions)
Wako kit, by Fujifilm (1 mentions)
Free glycerol reagent, by Merck Group (1 mentions)
Colorimetric assay kit, by Abcam (1 mentions)
Bradford assay, by Bio-Rad (1 mentions)
14 protocols using ffa quantification kit
1
Quantification of Brain FFA Content
2
Quantification of Free Fatty Acids
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The levels of FFAs in the extracts of the xenograft and the sera from mice were quantified in triplicate using the FFA quantification kit (Biovision, Mountain View, CA, USA) according to the manufacturer's instructions. Briefly, 10 mg of the xenografts were homogenized with 200 μL chloroform‐triton X‐100 (1% Triton X‐100 in pure chloroform), and the extracts were centrifuged at 500 rpm for 1 min. The purified lipids in the lower phase were collected, air dried and dissolved in 200 μL FA assay buffer. FAs were converted to their coenzyme A (CoA) derivatives, and the subsequent oxidized reaction and concomitant generated color were measured at 570 nm. The concentration was calculated using a PA standard curve, and the secreted FFA in the CM of the co‐culture plates of the cells was measured.
3
Blood Lipid and Glucose Measurement
4
Quantifying Liver and Plasma Lipids
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The levels of triglycerides in liver and plasma were measured with assay kits according to manufacturer’s instruction (Thermo Scientific, Waltham, MA). Concentrations of FFAs in liver and plasma were determined with a FFA quantification kit (BioVision, Milpitas, CA).
5
Quantifying Hepatic and Cardiac Lipid Profiles
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The hepatic and cardiac TC and TG contents were determined by Wako kits (Wako Pure Chemical Industries, Ltd.). For the measurement of free fatty acid (FFA) content, tissues were homogenized in 200 µl of chloroform with 1% Triton X‑100. Fatty acids were extracted in the chloroform fraction and N2-dried to remove the chloroform. Then, the hepatic and cardiac tissue FFA contents were determined by FFA quantification kit (BioVision, Mountain View, CA, USA)23 (link).
6
In vivo and ex vivo lipolysis assay
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To assess in vivo lipolysis, serum free fatty acids (FFAs) were measured in cardiac blood obtained from non-fasted mice before and 20 min after i.p. injection of CL316243 (1 mg/kg). Ex vivo and in vitro lipolysis was measured as described previously36 (link)50 (link) with some modifications. Briefly, for ex vivo analysis, adipose tissues were surgically removed from male and female mice, weighed, and washed with cold PBS; fat depots were incubated with CL316243 (1 μM) or vehicle in Krebs-Ringer Bicarbonate Buffer (KRBH, Sigma) containing 1% fatty acid-free BSA (GenDEPOT) and glucose (2.5 mM) for 2 h at 37 °C with mild shaking. After incubation, FFA and glycerol were measured using a glycerol-free reagent (Sigma) and the FFA quantification kit (BioVision, Mountain View, CA, USA) and normalized to the weight of adipose tissue samples.
7
Adipose Tissue Explant Acetaldehyde Assay
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Adipose tissue explants were obtained from the EWAT, cut into small pieces, and placed in a culture plate with pre-warmed PBS containing penicillin (100 U/ml) and streptomycin (100 mg/ml) to remove connective tissue and blood vessels. Explants (20 mg) were transferred to 24-well plates and cultured in 200 µl DMEM with L-glutamine (2 mM), penicillin (50 U/ml), streptomycin (50 mg/ml), and 2% fatty acid-free bovine serum albumin for 2 hours at 37 °C in the presence of absence of acetaldehyde (100 uM). FFA released in the culture medium was measured by a FFA Quantification Kit (BioVision, Mountain View, CA).
8
Quantifying Free Fatty Acids and Acetylcholinesterase in Mouse Retinae and Optic Nerves
9
Plasma and Myocardial Lipid Quantification
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Triglycerides (TG) and total cholesterol (TC) levels in the plasma were measured using an Olympus AU 600 auto analyzer (Olympus, Japan). Myocardial TG and TC were determined using an assay kit according to the manufacturer's instructions (Applygen, Beijing, China). The myocardial FFA content was determined with a free fatty acid (FFA) quantification kit (BioVision, San Francisco, CA).
10
Pituitary TG and FFA Quantification
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Pituitary TG was extracted and assayed in strict accordance with the kit manufacturer's instructions (Applygen Technologies Inc). Pituitary‐free fatty acids (FFAs) were assayed using an FFA quantification kit (BioVision). The contents of TG and FFAs were normalized to those of the corresponding proteins.
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