Name: Triglyceride Assay Kit - Quantification
SKU: ab65336
Rating: 4 (4 reviews)

Triglyceride Assay Kit ab65336 is a quantitative, addition-only assay with a 20 min and a 60 min incubation step. In the assay, triglycerides are converted to free fatty acids and glycerol, and the glycerol is then oxidized. Readout is on any colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) plate reader.

- Cited in over 500 publications
- Complete kit including standard curve for quantitation
- Used with sample types including tissue extracts, cell lysates, serum, heparin plasma, urine (UTI) and other biological fluids
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

Images

Functional Studies - Triglyceride Assay Kit - Quantification (AB65336), expandable thumbnail

Publications

Key facts

Detection method: Colorimetric/Fluorometric
Sample types: Heparin Plasma, Tissue Extracts, Serum, Other biological fluids, Cell Lysate, Urine (UTI)
Assay type: Quantitative
Assay time: 1h 20m
Sensitivity: > 2 µM

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Target data

See full target information Triglyceride

What's included?

2000 Test

Components

Assay Buffer 5

20 x 25 mL

Cholesterol Esterase

20 x 1 Vial

Enzyme Mix VI

20 x 1 Vial

OxiRed™ Probe

20 x 0.2 mL

Triglyceride Standard

20 x 300 µL

Key facts

Detection method: Colorimetric/Fluorometric
Sample types: Heparin Plasma, Tissue Extracts, Serum, Other biological fluids, Cell Lysate, Urine (UTI)
Assay type: Quantitative
Assay time: 1h 20m
Assay Platform: Microplate reader
Sensitivity: > 2 µM

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: -20°C
Appropriate long-term storage conditions: -20°C
Storage information: -20°C

Notes

Triglyceride Assay Kit (ab65336) is a sensitive, easy assay to measure triglyceride concentration in mammalian samples.

Other names used to refer to this type of assay include TAG assay, TG assay, and triacylglycerol assay.

How the Triglyceride assay works
In the triglyceride assay protocol, triglycerides are converted to free fatty acids and glycerol by a lipase enzyme. Glycerol is then phosphorylated by glycerol kinase, and subsequently oxidized by glycerol phosphate oxidase to produce hydrogen peroxide. The hydrogen peroxide reacts with a probe via a peroxidase to generate color (spectrophotometry at λ= 570 nm) and fluorescence (Ex/Em = 535/587 nm).

Triglyceride assay protocol summary

- Add samples and standards to wells.
- Add assay buffer and lipase, and incubate for 20 min.
- Add triglyceride reaction mix and incubate for 60 min.
- Analyze with microplate reader.

Getting the best performance from ab65336
Please note: the general range is 0-10 nmol (colorimetric) and 0-1 nmol (fluorometric).

If your sample contains reducing substances, they are likely to interfere with the assay. In this case, we recommend using Triglyceride Assay Kit (Fluorometric, Reducing samples) Triglyceride Assay Kit (Fluorometric, Reducing Samples) ab178780.

Technical Note: Triglyceride is unlikely to be detectable in healthy urine but may be elevated in the presence of infection or disease of the urinary tract.

How other researchers are using Triglyceride Assay Kit ab65336
The Triglyceride assay kit has been used in publications in a variety of sample types, including:

- Human: serum¹, plasma², mammary epithelial and mammary cancer cell line lysate³, Huh7.5 hepatocyte-derived cell line lysate⁴, primary liver cell line lysates⁵, sebocyte cell culture lysates⁶
- Mouse: hepatocyte cell lysates⁷, liver extract⁸, serum⁹, plasma¹⁰, kidney extracts¹¹, liver tissue and serum¹², cardiac tissue extracts¹³
- Rat: liver tissue extract¹⁴, plasma¹⁵
- Drosophila¹⁶

^{References: 1-Huang Y et al. 2019, PMID: 30778327, 2-Wilson et al 2018; PMID: 29534545, 3-Yen MC et al. 2019, PMID: 30911270, 4-Kim D et al. 2018, PMID: 30127285, 5-Boteon Y et al. 2018, PMID: 30044872, 6- Jin S and Lee MY 2018, PMID: 30400322, 7-Zhang W et al. 2019, PMID: 30664220, 8-Brial F et al. 2019, PMID: 30842494, 9-Liu J et al. 2018, PMID: 30344653 and Patton A et al. 2018, PMID: 29666152, 10-Fernandes et al. 2018, PMID: 29320745 and Körholz JC et al 2018, PMID: 30362941 and Liang et al. 2018, PMID: 29659562, 11-Ding W et al. 2018, PMID: 29563333, 12-Cui XB et al. 2018, PMID: 29449334, 13-Rohm M et al. 2018<. PMID: 29610263, 14-Yu S et al. 2018, PMID: 29867579, 15-García-Ruiz et al. 2018, PMID: 29367725, 16-Wen CA and Ballard JWO 2019, PMID: 30805163}

Related and recommended products
Triglyceride assay kit ab65336 is often used with Free Fatty Acid assay kit Free Fatty Acid Assay Kit - Quantification ab65341, and Cholesterol assay kit Cholesterol Assay Kit - HDL and LDL/VLDL ab65390, to study lipid metabolism and its role in various metabolic diseases such as diabetes, obesity, liver diseases, and cardiovascular conditions.

Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K622 Triglyceride Quantification Colorimetric/Fluorometric Kit. K622-100 is the same size as the 100 test size of ab65336.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Triglycerides also known as triacylglycerols are a type of lipid molecule found mainly in adipose tissue and the bloodstream. They consist of three fatty acids bonded to a glycerol backbone leading to a total molecular mass that varies based on the fatty acid chain lengths. Triglycerides originate in different tissues like the liver and intestines. The enzyme lipase acts on triglycerides breaking them down into free fatty acids and glycerol which is essential for metabolic processes. Scientists often measure triglyceride levels in blood to assess lipid metabolism and related health conditions.

Biological function summary

Triglycerides serve as a major form of energy storage in the body. They are important substrates for energy production especially during fasting or prolonged physical activity when the body's glycogen stores are depleted. Triglycerides do not function independently but as part of lipoprotein complexes. These complexes transport lipids throughout the body in the bloodstream. Understanding the normal triglyceride range helps in assessing if levels are within a healthy limit indicating effective lipid metabolism. Average triglyceride levels may vary but maintaining a good triglyceride level is important for overall health.

Pathways

Triglycerides are involved in lipid metabolism and energy homeostasis. One important pathway includes the lipolysis pathway where triglycerides undergo breakdown via lipase into fatty acids and glycerol. This reaction is important for providing energy particularly involving proteins such as hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL). In another pathway triglycerides play a role in the synthesis and storage of fats influenced by proteins like peroxisome proliferator-activated receptors (PPARs) which regulate gene expression involved in fatty acid storage.

Associated diseases and disorders

High triglyceride levels relate to cardiovascular disease and pancreatitis. Triglyceride-rich lipoproteins link to atherosclerosis making the regulation of triglycerides important for heart health. Chronic high levels may lead to acute pancreatitis by causing free fatty acid accumulation damaging pancreatic cells. Proteins such as apolipoprotein C-III affect triglyceride metabolism and may influence the risk of developing cardiovascular diseases. Effective management of triglyceride levels can help reduce these health risks.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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11 product images

Functional Studies - Triglyceride Assay Kit - Quantification (ab65336)
Triglyceride measured in cell culture lysates showing quantity (nmol) per 1 mln cells.
Samples with the concentration of 1e7 cells/mL were used. Samples were diluted 40-80 fold and measured fluorometrically.
HepG2 cells were treated with 25 uM Chloroquine for 72h.
Image from Samuel P et al., PLoS One 8(1), Fig 7b. doi: 10.1371/journal.pone.0048425. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Functional Studies - Triglyceride Assay Kit - Quantification (ab65336)
Hepatic triglyceride levels was measured using ab65336 in male and female wild-type (WT) or AT₂KO (knockout) mice with either normal diet (ND) or high fat diet (HFD).
Functional Studies - Triglyceride Assay Kit - Quantification (ab65336)
Fluorometric triglyceride standard curve: mean of duplicates (+/- SD) with background reads subtracted
Functional Studies - Triglyceride Assay Kit - Quantification (ab65336)
Colorimetric triglyceride standard curve using ab65336.
Biochemical assay - Triglyceride Assay Kit - Quantification (ab65336)
Triglyceride assay kit ab65336 and free fatty acid assay kit Free Fatty Acid Assay Kit - Quantification ab65341 used with heart tissue.
Han et al. used Triglyceride assay kit ab65336 and Free Fatty Acid assay kit Free Fatty Acid Assay Kit - Quantification ab65341 to investigate cardiac abnormalities in high fat diet fed mice when researching the role of the long non-coding RNA Lipid-Droplet Transporter (LIPTER).
The concentration of free fatty acids (FA) (f, n = 5 mice each group) were quantified using the Free Fatty Acid Assay kit (Free Fatty Acid Assay Kit - Quantification ab65341), and triglyceride (TG) concentrations (g, n = 7 mice each group) were quantified using the Triglyceride Quantification Assay Kit (ab65336).
Approximately 25 mg of mouse heart tissue per heart was collected and homogenized in 500 μl lipid extraction buffer (Lipid Extraction Kit (Chloroform Free) ab211044) using a Beadbug Homogenizer. After centrifugation, the supernatant was removed to a new tube. FA or TAG concentration was measured according to the manufacturer’s protocols. The FA and TAG concentrations in WT and genetically modified mouse hearts were normalized to the collected heart tissue weights.
Biochemical assay - Triglyceride Assay Kit - Quantification (ab65336)
Triglyceride assay kit ab65336 and Free Fatty Acid assay kit Free Fatty Acid Assay Kit - Quantification ab65341 used with mouse liver tissue.
Yue et al. used triglyceride assay kit ab65336 and free fatty acid assay kit Free Fatty Acid Assay Kit - Quantification ab65341 to investigate the protective effects of IL-22 on alleviating alcoholic hepatitis.
Administration of IL-22 reduces alcohol-induced lipid accumulation in mouse liver. PF (pair fed), AF (alcohol fed), AF+IL-22 (alcohol fed with IL-22 treatment).
(A) BODIPY 493/503 staining of mouse liver. Neutral lipids were stained in green, and nuclei were counterstained in blue. (B) Hepatic triglyceride (TG) and free fatty acid (FFA) levels.
Quantification assays of triglycerides (TG) and free fatty acids (FFA) in the liver were conducted using commercial kits from Abcam (Waltham, MA; ab65336 and Free Fatty Acid Assay Kit - Quantification ab65341) per the manufacturer’s instructions.
Biochemical assay - Triglyceride Assay Kit - Quantification (ab65336)
Diagram showing the principles of the Triglyceride assay method.
Biochemical assay - Triglyceride Assay Kit - Quantification (ab65336)
Triglyceride assay kit ab65336, Cholesterol assay kit Cholesterol Assay Kit - HDL and LDL/VLDL ab65390, and Free Fatty Acid assay kit Free Fatty Acid Assay Kit - Quantification ab65341 used with rat serum and rat liver.
Li et al. used Triglyceride assay kit ab65336, Cholesterol assay kit Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation ab65359, and Free Fatty Acid assay kit Free Fatty Acid Assay Kit - Quantification ab65341 to investigate the impact of using L. reuteri treatment to ameliorate the impact of biorhythm disorder-ignited dyslipidemia in long-term darkness treated rats.
The concentrations of triglycerides (TG), total cholesterol (CHOL), HDL-cholesterol (HDL-C), and free fatty acids (NEFA) in rat sera and liver tissues were detected using Triglyceride Quantification Assay Kit (ab65336), HDL and LDL/VLDL Cholesterol Assay Kit (Cholesterol Assay Kit - HDL and LDL/VLDL ab65390). and Free Fatty Acid Quantification Assay Kit (Free Fatty Acid Assay Kit - Quantification ab65341), respectively. All procedures were carried out following the recommended instructions provided by the kit manufacturers.
Biochemical assay - Triglyceride Assay Kit - Quantification (ab65336)
Triglyceride assay kit ab65336 and cholesterol assay kit Cholesterol Assay Kit - HDL and LDL/VLDL ab65390 used with mouse serum.
Zuo et al. used Triglyceride assay kit ab65336 and Cholesterol assay kit Cholesterol Assay Kit - HDL and LDL/VLDL ab65390 to investigate the impact of hepatocyte-specific AAV-mediated knockout of ANGPTL3 in mice.
Measurements of serum triglyceride (TG) and total cholesterol (TC) in mice at 1–4 weeks after AAV9-hAAT-Anc/Angptl3 administration. The control (Ctrl) mice received no treatments.
Blood samples collected from the animals were allowed for clotting at room temperature for 10 min. Serum was isolated by centrifugation at 5000 rpm for 10 min and stored at -80 °C in small aliquots. Determination of TG and TC was performed using the triglyceride and cholesterol assay kit (Abcam, ab65336 and Cholesterol Assay Kit - HDL and LDL/VLDL ab65390, respectively) according to the manufacture’s instruction.
Biochemical assay - Triglyceride Assay Kit - Quantification (ab65336)
Triglyceride assay kit ab65336 and free fatty acid assay kit Free Fatty Acid Assay Kit - Quantification ab65341 used with liver tissue and plasma samples respectively.
Samuel et al. used the triglyceride assay kit and free fatty acid assay kit to understand gender differences in metabolic syndrome disorders in angiotensin receptor AT2R knockout mice.
Plasma free fatty acid levels were measured by colorimetric method using the Abcam free fatty acid quantification kit.
Hepatic triglyceride levels was measured using ab65336 in male and female wild-type (WT) or AT2KO (knockout) mice with either normal diet (ND) or high fat diet (HFD). For hepatic triglyceride measurement, livers were dissected and snap-frozen in liquid nitrogen and stored at −80°C. In brief, lipids were extracted by homogenizing 25 mg liver tissue in 1 ml 5% Triton-X100 in water, then slowly heated to 80°C in water bath for 5 min. The samples were cooled down and again heated to solubilize all triglycerides into solution. The samples were centrifuged for 5 min and supernatants were diluted 10 fold with dH2O for quantification.
Biochemical assay - Triglyceride Assay Kit - Quantification (ab65336)
Triglyceride assay kit ab65336 and cholesterol assay kit Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation ab65359 used with HepG2 cell lysates.
Pozzi et al. used used the triglyceride assay kit and cholesterol assay kit to investigate the effect of antipsychotics on the SREBP pathway and on AMPK activation.
HepG2 cells serum-starved overnight and pre-treated with DMEM low glucose 20% FBS for 3 h, were incubated with 5 μM U18666A, 25 μM risperidone (RIS), ziprasidone (ZIP) or olanzapine (OLA), the SREBP2 positive control lovastatin 25 μM (LOV), and the SREBP1 positive control T0901317 10 μM (T09) for 24 h.
Triglycerides (TG) were extracted and quantified from HepG2 cells treated with the indicated compounds using Abcam ab65336. Briefly, 5 × 10^6 cells were harvested and lysed in 5% v/v NP-40, then samples were repeatedly heated at 98°C to solubilise triglycerides; a sample was taken for BCA protein quantification as normaliser, and the indicated amount of sample was processed with lipase and reaction reagents. Fluorescence emitted by the chromogen dye was quantified on Fluoroskan microplate reader (Ascent FL, Thermo Fisher Scientific) at 544/590 nm. Triglyceride fluorescence was reported as fold increase over levels of untreated cells (one way ANOVA followed by Dunnett’s multiple comparison test, n > 3 experiments; *vs. unt cells).
Total cholesterol was extracted from HepG2 cells treated with the indicated compounds by using cholesterol assay kit Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation ab65359. Briefly, 10^6 cells were harvested and lysed in 100 mM NaCl, 10 mM TRIS pH 7.4, 1 mM EGTA, 2 mM MgCl2 and 1% v/v Triton-X100; samples were repeatedly vortexed and chilled on ice to solubilise cholesterol; a sample was taken for BCA protein quantification as normaliser, while the rest was mixed with isopropanol-chloroform 11:7 and centrifuged to separate the organic phase containing cholesterol. The organic phase was taken and evaporated, cholesterol was then resuspended and processed with cholesterol esterase and reaction reagents. Fluorescence emitted by the chromogen dye was quantified on Fluoroskan microplate reader at 544/590 nm. Cholesterol fluorescence was reported as fold increase over levels of untreated cells (one way ANOVA followed by Dunnett’s multiple comparison test, n > 3 experiments; *vs. unt cells).