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Lipolysis (Adipocyte) Colorimetric/Fluorometric Assay Kit

Only kit on the market to isolate adipocytes from adipose tissue & measure lipolysis
Catalog #: K581

In stock

$520.00

Product Details

Cat # +Size K581-5
Size For 5 g Tissue
Detection Method Absorbance (OD 570 nm) and Fluorescence (Ex/Em = 535/587 nm)
Applications Isolation of adipocytes from rat/mouse tissue
Measurement of lipolysis by adipocytes
Screening compounds that influence lipolysis, mechanistic studies and studies on metabolic dysfunction
Features & Benefits • Simple, rapid & convenient method to isolate adipocytes from adipose tissue & measure lipolysis.
• The assay kit can detect less than 200 pmol glycerol in the colorimetric assay and less than 20 pmol in the fluorometric assay.
Kit Components • Collagenase (0.2%)
• Collagenase Stop Buffer
• Adipocyte Wash Buffer
• Adipocyte Lipolysis Buffer
• Glycerol Assay Buffer
• Glycerol Probe (in DMSO, anhydrous)
• Glycerol Enzyme Mix (Lyophilized)
• Glycerol Standard (100 mM)
• Isoproterenol (10 mM)
• Cell strainer (100 µm)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Lipolysis is the intracellular hydrolysis of triglycerides into glycerol and free fatty acids, which are then released into the bloodstream or culture media. Lipolysis occurs in essentially all cells, but is most abundant in white and brown adipose tissue. Deficiencies in lipolysis lead to increased intracellular lipid accumulation, resulting in abnormal cellular physiology, hyperlipidemia, and insulin resistance. Lipolysis can be induced by hormones or catecholamine, which binds to β-adrenergic receptors leading to activation of adenylate cyclase, forming cAMP from ATP. cAMP serves as a second messenger to activate hormone-sensitive lipase, which hydrolyzes the triglycerides. This pathway is inhibited by insulin. BioVision’s Adipocyte Lipolysis Colorimetric/Fluorometric Assay kit is a simple, facile way to monitor lipolysis. The kit provides all reagents necessary to isolate adipocytes from up to 5 g of mouse or rat adipose tissue and measure glycerol release from primary adipocytes after induction of lipolysis. It also includes the synthetic catecholamine, Isoproterenol, to stimulate the cAMP-mediated pathway. The signal intensity is directly proportional to the amount of glycerol present. This assay kit can detect less than 200 pmol glycerol in the colorimetric assay and less than 20 pmol in the fluorometric assay.


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What do you mean with "collect medium" after the lipolysis stimulation? Do i have to centrifuge the aliqouts after stimulation and take the bottom of the solution, or the top layer?
Suspend the adipocyte pellet in the Lipolysis buffer and aliquot in different tubes. Incubate them with and without isoproterenol. After the stimulation collect the medium, centrifuge and take the supernatant from the top for the assay.
Am I suppose to remove the wash buffer from the bottom of the solution? Or from the top?
Remove and discard the supernatant from the top.
Can you skip the collagenase digestion step?
Adipose cell cluster size is reduced by collagenase digestion, while eliminating contaminating proteins and blood residues from the final cell product suspension. http://www.hindawi.com/journals/bmri/2012/462543/
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. You have 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.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
 BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.