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Product Details
| Cat # +Size | K630-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm) |
| Applications | The kit can detect 50 pmol-10 nmol (or 1~10000 µM range) of glycerol in various samples. |
| Features & Benefits | • Simple procedure; takes ~ 40 minutes • Fast and convenient • The kit provides an accurate assay for measuring free glycerol in various samples |
| Kit Components | • Glycerol Assay Buffer • Glycerol Probe (in DMSO, anhydrous) • Glycerol Enzyme Mix (lyophilized) • Glycerol Standard (100 mM) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Glycerol is widely used in foods, beverages, solvents, pharmaceutical and cosmetic products, etc. There is broad interest in quantification of glycerol for research and development. BioVision’s Glycerol Assay Kit provides a sensitive, easy assay to measure free glycerol concentration in various samples. In the assay, glycerol is enzymatically oxidized to generate a product which reacts with the probe to generate color (λ= 570 nm) and fluorescence (Ex/Em = 535/587 nm). The kit can detect 50 pmol-10 nmol (or 1~10000 µM range) of glycerol in various samples.
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How can cells/tissue samples be used with this kit?
Start with 1-2 million cells (10-100mg tissue), suspend the cell pellet in 500 µl (or ~4 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Load the supernatant unto a 10kda spin column for deproteinzation. Use the eluate for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.
Can this kit be used with bacterial/yeast samples?
All BioVision kits are optimized with mammalian samples but can work with many other species including yeast. One consideration is that yeast cells have cell walls and hence might need specific reagents to break the cell wall in yeast cells and some bacterial cells.
Can I use the same method of samples preparation in the protocol of Triglyceride Quantification Kit to prepare the samples to use this Free Glycerol Assay Kit to measure intracellular glycerol?
This will not work. For the triglyceride assay lipid extracts are prepared from the cells while for the Free glycerol assay cell lysate/tissue homogenate is used.
Should medium to be assayed by this kit be stored at -20C or 4C?
We recommend storing the medium at -20C or preferably at -80C for best results. If the assay will be done in multiple batches, aliquots can be stored to minimize freezing-thawing.
What is the detection range for the colorimetric assay?
The colorimetric assay shows a linear standard curve in the micromolar -mM concentration range of glycerol.
Can glycerine be used in this assay?
Yes, Glycerine is same as glycerol and can be used with this kit.
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 customized dilutions be made for the std curve to fit the expected range for the sample data?
Yes, the standard (100mM) can be diluted to prepare a std. curve to suit the requirement of the sample type.
Why are the standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, the incubation time can be increased. As long as the standard curve is linear, it should be fine to use, since all the samples will also be measured under the same conditions on this curve.
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.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. It is recommended to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Iso, T. et al., (2017) Serum levels of fatty acid binding protein 4 and fat metabolic markers in relation to catecholamines following exercise, Clinical Biochemistry, Jun.2017, https://doiorg/101016/jclinbiochem201705021
Tsao et al, Interleukin-4 regulates lipid metabolism by inhibiting adipogenesis and promoting lipolysis. J. Lipid Res., Mar 2014; 55: 385 - 397.
MacDonald et al., IL-6 and epinephrine have divergent fiber type effects on intramuscular lipolysis. J Appl Physiol, Nov 2013; 115: 1457 - 1463.
Tetsuro Izumi et al., Activin Receptor-Like Kinase 7 Suppresses Lipolysis to Accumulate Fat in Obesity Through Downregulation of Peroxisome Proliferator–Activated Receptor and C/EBPα. Diabetes, Jan 2013; 62: 115 - 123.
Özlem Gögebakan et al., Glucose-Dependent Insulinotropic Polypeptide Reduces Fat-Specific Expression and Activity of 11β-Hydroxysteroid Dehydrogenase Type 1 and Inhibits Release of Free Fatty Acids, Diabetes, Feb 2012; 61: 292 - 300.
For more citations of this product click here
