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Adipogenesis Colorimetric/Fluorometric Assay Kit

based on 7 citations in multiple journalsAdipogenesis Colorimetric/Fluorometric Assay Kit74.2 4
Highly Sensitive Assay
Catalog #: K610

In stock

$325.00

Product Details

Cat # +Size K610-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The kit can detect triglyceride in as few as a thousand or less differentiated 3T3-L1 cells with triglyceride detection linear range 0.2 to 10 nmol.
Features & Benefits • Simple procedure; takes ~40 minutes
• Fast and convenient
• Kit contains all necessary reagents for rapid, sensitive and accurate measurement of Adipogenesis in various samples.
Kit Components • Adipogenesis Assay Buffer
• Lipid Extraction Solution
• Adipogenesis Probe (in DMSO solution)
• Lipase (lyophilized)
• Adipogenesis Enzyme 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

Adipogenesis is the process of differentiation of different cell types into adipocytes, the primary fat storage cell type. The accumulation of adipocytes is the basis for obesity, a significant risk factor in many diseases, including diabetes, atherosclerosis, cancer and cardiovascular disease, etc. Adipocytes accumulate triglycerides, in the form of lipid droplets which can be measured. BioVision’s adipogenesis assay kit quantifies triglyceride accumulation in cells and tissues. In the assay, triglycerides are efficiently solubilized then hydrolyzed to glycerol which is subsequently oxidized to convert the probe to generate color (λmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The kit can detect triglyceride in as few as a thousand or less differentiated 3T3-L1 cells with triglyceride detection linear range 0.2 to 10 nmol. The high detection sensitivity and the convenient microplate assay format make the kit a convenient tool for studying the effect of adipogenesis inducers or inhibitors, or to screen drugs.


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Can protein content be used as an internal control for this assay?
Yes, a detergent-compatible BCA assay can be used for protein quantitation to normalize sample amount.
Will the Assay buffer help in cell lysis? IS it necessary to prepare a cell lysate before proceeding with the protocol?
There is no need to prepare a cell lysate. The Lipid extraction solution can be used according to the protorol for this purpose. For tissues, on the other hand, flash frozen, pulverized tissue can be used for lipid extraction. Otherwise, minced fresh tissue can be homogenized in the lipid extraction solution before heating the sample for lipid extraction.
Can cholesterol be assayed using this kit?
For measuring cholesterol in samples cholesterol assay kits K623 (colorimetric) or K603 (colorimetric/fluorometric) can be used. K^10 measures totl lipids in a sample.
Is this kit appropriate for measuring lipids in blood or other body fluids?
The adipogenesis kit is usually used with adipocytes or adipose tissue, hardly with body fluids since there is not much adipogenesis going on in the fluids. Larger volume of sample might be required to test aqueous body fluids with this kit.
Can this kit be used for testing lipid accumulation in hepatocytes from fatty liver?
Yes, liver tissue can be homogenized in the lipid extraction solution and the lipids can be assayed for.
Will all cells follow the same differentiation/induction pattern as shown for 3T3 cells on the datasheet?
No, this is not necessary. Optimization of the induction of adipogenesis and the differentiation time course must be tested and validated before using cells with this assay.
What is the best way to store samples to be assayed with this kit?
" Cells can be frozen at -80C in a small volume of the adipogenesis assay buffer. The lipid extraction solution might lyse cells immediately and that might affect the results after storage. On the day of the experiment, remove the assay buffer and then add the lipid extraction solution to proceed with the assay. The other option is to extract the lipids and then store the extract at -80C to be assayed at a later date. We would prefer the first option though to prevent lipid aggregation and other issues like lipid extract sticking to the tube walls. The other option is to extract the lipids and then store the extract at -80C to be assayed at a later date. We would prefer the first option though to prevent lipid aggregation and other issues like lipid extract sticking to the tube walls."
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.
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.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
What is the exact volume of sample required for this assay?
The amount of the lipid extract used for this assay will depend on cell type, treatment and cell differentiation stage. For unknown samples, we suggest testing different volumes of a few representative samples to make sure the readings are within the standard curve linear range. used for the triglyceride assay will depend on celltype, treatment and cell differentiation stage.For unknown samples, we suggest testing different doses of your sample to make sure thereadings are within the standard curve range.
Haffar et al., Saturated fatty acids induce endoplasmic reticulum stress in primary cardiomyocytes. Endoplasm. Reticul. Stress Dis. 2015; 2, 53–66.
Abdulla et al., Regulation of Lipogenic Gene Expression by Lysine-specific Histone Demethylase-1 (LSD1). J. Biol. Chem., Oct 2014; 289: 29937 - 29947.
Madak-Erdogan et al., Integrative genomics of gene and metabolic regulation by estrogen receptors α and β, and their coregulators. Mol Syst Biol, Jul 2014; 9: 676.
McIntosh et al., Human FABP1 T94A variant impacts fatty acid metabolism and PPAR-α activation in cultured human female hepatocytes. Am J Physiol Gastrointest Liver Physiol, Jul 2014; 307: G164 - G176.
Aung Than et al., Control of Adipogenesis by the Autocrine Interplays between Angiotensin 1–7/Mas Receptor and Angiotensin II/AT1 Receptor Signaling Pathways. J. Biol. Chem., May 2013; 288: 15520 - 15531.
For more citations of this product click here