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Triglyceride Quantification Colorimetric/Fluorometric Kit

based on 68 citations in multiple journalsTriglyceride Quantification Colorimetric/Fluorometric Kit684.7 5
Catalog #: K622

In stock

$485.00

Product Details

Cat # +Size K622-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The kit can detect 2 pmol-10nmol (or 2-10000µM range) of triglyceride in various samples. The kit also detects monoglycerides and diglycerides.
Features & Benefits • Simple procedure; takes ~1 hour
• Fast and convenient
• Kit contains the necessary reagents for accurate measurement of triglyceride in various biological samples
Kit Components • Triglyceride Assay Buffer
• TriglycerideProbe (in DMSO, anhydrous)
• Lipase
• TriglycerideEnzyme Mix (lyophilized)
• Triglyceride Standard (1 mM)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Triglycerides (TG) are the main constituent of vegetable oil, animal fat, LDL and VLDL, and play an important role as transporters of fatty acids as well as serving as an energy source. TG are broken down into fatty acids and glycerol, after which both can serve as substrates for energy producing and metabolic pathways. High blood levels of TG are implicated in atherosclerosis, heart disease and stroke as well as in pancreatitis.The Triglyceride Quantification Kit provides a sensitive, easy assay to measure TG concentration in a variety of samples. In the assay, TG are converted to free fatty acids and glycerol. Theglycerol is then oxidized to generate a product which reacts with theprobe to generate color (spectrophotometry at λ = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The kit can detect 2 pmol-10nmol (or 2-10000µM range) of triglyceride in various samples. The kit also detects monoglycerides and diglycerides.


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How specific this kit is for measuring triglycerides?
The lipase generates glycerol – as well as other products from phospholipids. Glycerol kinase phosphorelates glycerol which is further reduced to develop color. Glucokinase specificity is very high as is the further enzymatic processing, which will not react with phospholipids. When we used phosphatidyl choline, for example, we did not get any color showing that the kit is very specific.
What can be done if the lysed cells are not dissolving?
The amount of 5 % NP-40 in water used can be increased. Also, the temperature can be reaised above 80° C to get the particles into solution in addition to repeating the heating and cooling for 2 cycles.
What could be the explanation for negative OD values but positive BODIPY staining in cells?
The kit can detect 2 pmol -10nmol (or 2 – 10000 uM range) of triglyceride in various samples. It could be that the BODIPY staining is showing total lipids in these cells but the amount of triglycerides is low. The fluorometric version of this assay using the same kit is 10x more sensitive than the colorimetric one and could be chosen to see if readings make sense. Also, it is crucial to check the instrument settings while measuring the samples. Use the correct filter for 570nm detection.
How much Triglyceride is there is serum samples?
Typically serum levels of Triglyceride can range between 0.1-6mM, individual experimental findings may vary.
How do we convert mM to mg/ml?
"A solution strength of 1 mol/L is represented as 1mM, so for example, 50 mM is 50 mmol/L (millimoles per liter). Divide by the molecular weight of the substance (average triglyceride molecular weight in your sample) to convert from moles to grams. Then mM/mol weight can be converted to get mg/L. For this assay, since it is difficult to say the molar mass of triglyceride, unless which specific one is present in the sample is known, the accepted most common way of reporting the data is as mM or µmol/ml."
Is it generally possible to maintain the samples in NP-40/water at -20C and measure their triglyceride concentration again? Additionally, are the triglycerides concentrated on the surface layer of the supernatant or distributed in the whole supernatant after the boiling step?
It is possible that triglycerides aggregate upon freezing and thawing while in an aqueous solution and stick to the walls of the tube which can skew the results. There could be a layer of fat/oil after boiling. Once the sample is centrifuged, the liquid can be collected in a fresh tube and thoroughly vortexed so that when the sample is added to a well, a homogenous solution is pipetted.
Can lesser than 10 million cells be used for this assay?
Less cells can be used, but the yield of triglycerides might be less. The number of cells depends on the amount of triglycerides in them. If less cells are used the volume of NP40-water can be scaled down proportionately.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Zhang W., et al., (2018) , Adipose-specific lipin1 overexpression in mice protects against alcohol-induced liver injury, Sci. Rep., 2018, 8:408
Sundaram, M. et al., (2017) he apolipoprotein C-III (Gln38Lys) variant associated with human hypertriglyceridemia is a gain-of-function mutation, Journal of Lipid Research, Sep.2017, jlr-M077313
Zhang P et al., (2017) Beraprost sodium, a prostacyclin analogue, reduces fructose-induced hepatocellular steatosis in mice and in vitro via the microRNA-200a and SIRT1 signaling pathway, Metabolism, 2017, 73:9-21
Tse, Margaret Chui Ling et al. (2017) Tumor Necrosis Factor a Promotes Phosphoinositide 3-Kinase Enhancer A and Amp-Activated Protein Kinase Interaction to Suppress Lipid Oxidation in Skeletal Muscle, Diabetes. 2017 Jul;66(7):1858-1870.
Horwath, Julie A. et al. (2017) Obesity-induced hepatic steatosis is mediated by endoplasmic reticulum stress in the subfornical organ of the brain, JCI Insight. 2017 Apr 20;2(8).
Shimazu, Hiroshi et al. (2017) Pharmacological targeting of plasmin prevents lethality in a murine model of macrophage activation syndrome, Blood. 2017 Mar 21.
For more citations of this product click here
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