Lipolysis Isolation and Quantification Assay Kit (Colorimetric/Fluorometric) (ab196987)

Overview

  • Product name

    Lipolysis Isolation and Quantification Assay Kit (Colorimetric/Fluorometric)
    See all Lipolysis kits
  • Detection method

    Colorimetric/Fluorometric
  • Sample type

    Tissue
  • Assay type

    Quantitative
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Lipolysis Isolation and Quantification Assay Kit (Colorimetric/Fluorometric) (ab196987) is a simple assay to monitor lipolysis using primary adipocytes. The kit provides all reagents necessary to isolate up to 5 grams of adipocytes from mouse or rat adipose tissue and measure glycerol release after induction of lipolysis. It also includes a synthetic catecholamine, isoproterenol, which stimulates the cAMP-mediated lipase activation pathway. The signal intensity is directly proportional to the amount of glycerol present.


    This product can detect from 20 pmol glycerol when using the fluorometric assay or from 200 pmol glycerol when using the colorimetric assay.

  • Notes

    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 such as 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.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 5 g
    Adipoctye Wash Buffer 1 x 22ml
    Adipocyte Lipolysis Buffer 1 x 17ml
    Cell Strainer (100 μm) 10 units
    Collagenase 1 x 10ml
    Collagenase Stop Buffer NM 1 x 90ml
    Glycerol Assay Buffer 1 x 25ml
    Glycerol Enzyme Mix (lyophilized) Green 1 vial
    Glycerol Probe 1 x 0.2ml
    Glycerol Standard (100 mM) 1 x 0.2ml
    Isoproterenol (10mM) 1 x 50ml

Images

  • Measurement of glycerol level from adipocyte samples (20 µL of 1/10 diluted sample) using fluorometric assay. Adipocytes were either treated with vehicle control (H2O) or 100 nM isoproterenol for 0-3 hours.

  • Glycerol Standard Curve example for fluorometric assay.

  • Glycerol Standard Curve example for colorimetric assay.

Protocols

References

This product has been referenced in:

  • García-Ruiz I  et al. Omentectomy Prevents Metabolic Syndrome By Reducing Appetite and Body Weight In A Diet-Induced Obesity Rat Model. Sci Rep 8:1540 (2018). WB . Read more (PubMed: 29367725) »
See 1 Publication for this product

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