Navigation Links
FREE FAT CELL ASSAY

Susanne Rasmussen, Anders R. Srensen Novo Nordisk A/S

Introduction

The free fat cell assay has been used extensively to demonstrate the effect of insulin and insulin analogues on target tissue (1,2). The principle of the assay has been described by Rodbell(3) and Moody et al(4). The assay uses isolated adipocytes from either mice or rats. When stimulated by insulin, the adipocytes incorporate glucose into fat depots, and a suitably labelled glucose added to the medium will result in incorporation of radioactivity into the lipid pool. Extraction of the lipids from the cells has traditionally been performed using a toluene-based scintillator. The phase separation in this system is so effective that direct liquid scintillation counting of the sample is possible without further preparation. The counts originating from the aqueous phase, which still contains a considerable amount of radio labelled glucose, are negligible. Previously, the scintillators that were available for use in the TopCount microplate scintillation and luminescence counter did not have the preferred characteristics for the phase separation, so the toluene scintillator method, with its many undesirable chemical and physical properties, continued to be used. However, the availability of MicroScint-E scintillator from Packard Instrument Company now makes it possible to discontinue the use of toluene-based scintillator solutions, and count samples prepared with MicroScint- E directly in the TopCount.

Method

Animals are killed, and the epididymal fat pads are removed and placed in degradation buffer. Degradation is carried out at 37 C under vigorous shaking for one hour. The cell suspension is filtered in order to remove tissue debris, and the adipocytes are washed twice and resuspended in incubation buffer.

Using the 96-well microplate format, 100 μL of cell suspensio
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. DELFIA Protein Kinase assays.
2. A Positive Selection Assay for Mutation Analysis in Big Blue Animals
3. Luminol-enhanced Assay for Superoxide Anion (O2 face="Symbol" )
4. Organize Your Data from the Big Blue Mutation Assay
5. Fluorescence-Based Single-Tube Assays to Rapidly Detect Human Gene Mutations
6. RNase Protection Assay (RPA) Using DIG-Labeled RNA Probes
7. measuring cell proliferation with SpectraMax M5, M2 and Gemini microplate readers using the CyQUANT Cell Proliferation Assay
8. Matched siRNAs and Assays: Ambion + Applied Biosystems = RNAi Success
9. Using Validated siRNAs in Functional Genomic Assays
10. Nuclease Protection Assays: The Basics
11. RNArticles: RNA Amplification Used to Develop a New In Vitro HCMV Assay
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: FREE FAT CELL ASSAY

(Date:7/10/2014)... OMICS Group’s 5th International conference and ... 2014 at Double Tree by Hilton Beijing, China is ... Analytical & Bioanalytical research methods only to facilitate improved ... occasion, Dr. Srinubabu Gedela, MD of OMICS Group Inc ... remarkable one in bringing a unique and international mix ...
(Date:7/10/2014)... POINT, Ind. , July 10, 2014 ... expanded its use of capnography for respiratory ... growing group of healthcare leaders in embracing ... measures how effectively patients are breathing and ... depression occurs. By measuring the amount of ...
(Date:7/10/2014)... Utah , July 10, 2014 /PRNewswire-iReach/ ... annotation and interpretation, today announced an agreement ... (NGS) analytics for Lineagen,s NextStep Dx ... of neurodevelopmental and neurological disorders, currently offers ... microarray (CMA) testing service used by healthcare ...
(Date:7/10/2014)... to the International Atomic Energy Agency (IAEA) the ... acquiring sufficient quantities of plutonium or highly enriched ... device. The IAEA also notes that most cases ... which can be challenging to detect with most ... appearing this week in the Journal of ...
Breaking Biology Technology:A Critical Review of Analytical Techniques for Improved Living Standards 2A Critical Review of Analytical Techniques for Improved Living Standards 3A Critical Review of Analytical Techniques for Improved Living Standards 4Franciscan St. Anthony Health-Crown Point Underscores Patient Safety Commitment Through Expanded Use of Capnography 2Tute Genomics Platform Selected to Provide Clinical Interpretation for Lineagen's NextStepDx PLUS Next-Generation Sequencing-Based Diagnostic Test 2Tute Genomics Platform Selected to Provide Clinical Interpretation for Lineagen's NextStepDx PLUS Next-Generation Sequencing-Based Diagnostic Test 3Tute Genomics Platform Selected to Provide Clinical Interpretation for Lineagen's NextStepDx PLUS Next-Generation Sequencing-Based Diagnostic Test 4Sensitive detection method may help impede illicit nuclear trafficking 2
... , DURHAM, N.C., Nov. 27 Cromoz ... nanotube-based cancer drug delivery system in Hyderabad, India. The water-soluble ... allows for conjugation with cancer drugs, was developed in partnership ... nanotubes serves as a drug delivery vehicle with the ability ...
... of ultrasmall transistors and more powerful computer chips using ... after a key discovery by researchers at IBM, Purdue ... The researchers have learned how to create nanowires ... at the atomic level, which is a critical requirement ...
... ... of Canada , ... (Vocus) November 25, 2009 -- Ambassador to Speak at Economic Club of Canada., , , ... Invest in France Agency ,To view a biography of David Appia go to www.investinfrance.org ...
Cached Biology Technology:North Carolina Based Cromoz Inc. Launches Carbon Nanotechnology for Target Drug Delivery System in Cancer Treatment in Hyderabad, India 2Nanowires key to future transistors, electronics 2Nanowires key to future transistors, electronics 3French Ambassador for International Investment to Visit Toronto December 11 2
(Date:7/11/2014)... -- Research and Markets has announced the ... Industry Analysis Size Share Growth Trends and Forecast to ... http://photos.prnewswire.com/prnh/20130307/600769 The rising number ... for high level security in both private and public ... username and passwords, tokens etc. are vulnerable to attacks. ...
(Date:7/11/2014)... de Janeiro, Brazil- In the brains of all ... that allows an electric or chemical signal to ... Chemical synapses, which are the most abundant type ... Synapse formation is crucial for learning, memory, perception ... inhibitory synapses critical for brain function. For ...
(Date:7/10/2014)... , June 27, 2014  The American Academy ... Institute of Standards and Technology (NIST) and the ... academic research and forensic science expert members to ... is an element of the NIST,s Organization of ... among the first appointments made to the new ...
Breaking Biology News(10 mins):International Biometrics Technology Market - Industry Analysis Size Share Growth Trends and Forecast to 2019 2Blame it on the astrocytes 2Ten Members of the American Academy of Forensic Sciences Appointed to Forensic Science Standards Board (FSSB) 2Ten Members of the American Academy of Forensic Sciences Appointed to Forensic Science Standards Board (FSSB) 3
... reduces spoilage explains a scientist at the Society for ... explains how ozone treatment could be a safe, effective ... foods. It is estimated that up to 30% ... spoilage. Dr Singleton from Newcastle University explains that low ...
... being able to develop a computer model of the brain ... functions of nerve cells in the brain together for the ... in the neuroscience known as ,connectomics,. With parallels to genomics, ... the brain,s connections (known as ,synapses,). By mapping these connections ...
... Lee, a graduate student in Cornell University,s Dyson School of ... known as "the halo effect" for some time. Psychologists have ... of a person can be influenced by how we perceive ... the fact that a person has a positive attribute can ...
Cached Biology News:Ozone reduces fungal spoilage of fruits and vegetables 2Mapping the brain: New technique poised to untangle the complexity of the brain 2Mapping the brain: New technique poised to untangle the complexity of the brain 3The health halo effect: Don't judge a food by its organic label 2
... Provides up to 30 minutes of walkaway time. Automatic Calibration. Five minute cooldown ... port. ... ... ...
Vybrant® Multidrug Resistance Assay Kit...
...
... culture systems provide an integrated set-up ... and imaging. Each system uses medical-grade ... coverslips in cell culture plates. The ... ready to use. Individual chambered coverslips ...
Biology Products: