Friday, September 28, 2012

Focus On a Fluorescent Alternative Assay to the Caco-2 Study for Drug Membrane Permeability

New technologies and methods are continuously created that supplant outdated methods in order to save time, money, and to be safer. Now with the PHERAstar FS microplate reader, there are safer, faster, and more cost-effective alternatives to some popular radioactive assays.


One such assay is the Fluorosome trans-PGP application that can replace the Caco-2 assay. This assay measures membrane permeability through the action of P-glycoprotein (PGP) pump. Using purified PGP protein in liposomes, this assay occurs in minutes rather than days and is specific for just the PGP protein.


For more information see the applications noteA faster, more reliable alternative is the Fluorosome®-trans-PGP assay from the Fluorosome Company

Thursday, September 27, 2012

Applications Thursday: New Reference Article using the PHERAstar for fluorescence anisotropy measurements

In a recently published article in the Journal of biomolcular Screening, the PHERAstar equipped with a fluorescence polarization Optic Module was used for anisotropy measurements:


A High-Throughput–Compatible Fluorescence Anisotropy-Based Assay for Competitive Inhibitors of  Escherichia coli UDP-N-Acetylglucosamine Acyltransferase (LpxA)


The article was written by Adam B. Shapiro, Philip L. Ross, Ning Gao, Stephania Livchak,
Gunther Kern, Wei Yang, Beth Andrews, and Jason Thresher from AstraZeneca R&D Boston, Waltham, MA, USA.

You can find an abstract of the publication here:

http://jbx.sagepub.com/content/early/2012/09/25/1087057112462062.abstract

For more information on fluorescence polarization and a list of BMG LABTECH instruments that an perform fluorescence polarization measurements visit: http://www.bmglabtech.com/technology/fluorescence-polarization.cfm


Wednesday, September 26, 2012

Fun Fact: The Ig Nobel Prizes - For Research that Makes You Laugh and Think

The 22nd First Annual Ig Nobel Prizes were recently awarded, just weeks before the actual Nobel prizes are announced. The Ig Nobel Prizes, as explained, “honor achievements that first make people laugh, and then make them think. The prizes are intended to celebrate the unusual, honor the imaginative — and spur people's interest in science, medicine, and technology.”  The magazine Annals of Improbable Research organizes the Ig Nobel Prizes each year, with the annual ceremony held as the Harvard Sanders Theatre.
Ten  prizes are given out and this year’s 2012 Ig Nobel Prizes winners include:

Tuesday, September 25, 2012

Did you know that BMG LABTECH will be at Chemical Biology 2012 in Heidelberg, Germany, September 26-29?

BMG LABTECH is a proud, local sponsor of the Chemical Biology 2012 biennial EMBO conference in Heidelberg, Germany. This is the third Chemical Biology conference to occur in the Advanced Training Centre at EMBL Heidelberg, with the event getting bigger each year. The popularity of the conference stems from the growing need for interdisciplinary research - Chemical Biology emphasizes the use of chemical tools and strategies to answer biological questions. Amongst the topics covered, include:

  • Molecular imaging and switching tools
  • Computational and structural approaches
  • Innovative drug design approaches
  • Chemical genetics and high throughput methods
  • Novel synthetic methods as applied to biological questions
  • Chemical protein engineering
  • Chemical cell biology

EMBL building, HeidelbergBMG LABTECH will be highlighting the SPECTROstar Nano absorbance microplate reader and the PHERAstar FS HTS microplate reader. Both instruments have BMG LABTECH’s unique spectrometer-based absorbance, which allows for ultra-fast, full spectrum (220-1000 nm) measurements. These spectrometer-based microplate readers are perfect for full spectrum applications, such as the SoPRano™ Label-free protein-protein binding assay, which will be highlighted in a poster.

For more information see: http://www.embl.de/training/events/2012/CHB12-01/.

Monday, September 24, 2012

FAQ: Is there a fluorescence alternative to the hERG channel patch clamp assay?

Competitive binding assay of 10 known
hERG blockers using the
Predictor™ hERG Fluorescence Polarization Assay

The human ether-a-go-go related gene (hERG) potassium channel has been shown to be inhibited by certain drugs, which can cause cardiotoxic side effects such as long Q-T syndrome. Therefore all new drugs have to be tested for interaction with the hERG channel. The classical way to test the hERG channel is with patch clamp studies, but this is time consuming and it requires specialized electrophysiological equipment.

Now there is a more cost-effective, faster alternative to the hERG patch clamp assay that is used to test drug-induced long Q-T syndrome. It is the Predictor® hERG cellular FP assay from Life Technologies. This fluorescence polarization application saves money and time by allowing potential drugs to be prescreened before undergoing more tedious patch clamp screening.

Find more information here: Application Note 173: Predictor™ hERG Fluorescence Polarization Assay Kit performed on the PHERAstar/PHERAstar Plus

Friday, September 21, 2012

Focus On: Lab Manager’s Product Focus: Microplate Readers


Angelo DePalma recently published a product focus for Lab Manager on microplate readers. Dr. E.J. Dell from BMG LABTECH is quoted, as well as several other experts in the field from differing companies. Their views and thoughts are captured in this article, including upcoming trends in microplate reader technology and the things to consider when purchasing a new microplate reader.

Here is an excerpt:
According to Dell, during the past five years microplate readers have undergone “major advances that have broadened their use” and have “transitioned from being a luxury item in a well-financed facility to being a mandatory piece of equipment for a new start-up laboratory.” BMG LABTECH has introduced technologic advances related to absorbance detection and bottom reading (required for cell-based assays).
Read more from the article here: http://www.labmanager.com/?articles.view/articleNo/30217/

Thursday, September 20, 2012

Applications Thursday: Label-free Protein-Protein Binding Kinetics Using SoPRano(TM) Gold Nano Rods

Pharma Diagnostics’ standard SoPRano™ Gold Nano Rod (GNR) kits now allow KD, ka and kd to be simply and rapidly derived without the need for expensive dedicated SPR instrument. The new SoPRano™ kinetics application is enabled exclusively on BMG LABTECH spectrometer-based absorbance readers via a dedicated software upgrade, uniquely delivering real-time, solution phase kinetics for protein-protein interactions in cuvettes or microtitre plates (96 and 384-well).

As previously reported, Pharma Diagnostics’ SoPRano™ label-free platform has been certified to work on all BMG LABTECH microplate readers that have a CCD spectrometer, delivering high-quality, automated label-free analysis for end-point assays, IC/EC50 and KD determination for protein-protein interactions (Application Note 230). The addition of the new SoPRano™ kinetics application and software, now adds real-time kinetics data generation ability, thereby delivering a powerful and relatively inexpensive tool for all aspects of protein-protein interaction research.

For more information see Application Note 231: Label-Free Kinetic Measurements Using SoPRano™ Gold Nano Rods (GNRs) and a Spectrometer Microplate Reader.

For more information on SoPRano™ please go to www.pharmadiagnostics.com.

Wednesday, September 19, 2012

Fun Fact: September 19, 2012


Squamous cell carcinoma in the cervix, pap stain Born today in 1915, was Elizabeth Stern Shankman. She was a Canadian pathologist who was one of the first researchers to specialize in cytopathology, or the study of diseased cells. She performed breakthrough research in cervical cancer, having demonstrated that a normal cell undergoes 250 distinct stages before reaching an advanced cancerous state. Her research allowed for earlier diagnostic testing for cervical cancer, thereby greatly reducing the fatality rate of that disease.

She is also recognized as the first researcher to show that a specific cancer (cervical) is linked to a specific virus (herpes simplex). In addition, she was the first to show a link between prolonged use of oral contraceptive pills and cervical dysplasia. She died in 1980 at the age of 64 from stomach cancer.

Elizabeth Stern Shankman has also been named one of the 300 women who have changed the world, according to Encyclopaedia Britannica.

Tuesday, September 18, 2012

Did you know that the MARS Data Analysis software has a hyperbola curve fit that can be used for protein-protein interactions?

After defining standards, a hyperbola curve fit can be used to create a standard curve in the MARS Data Analysis software. A hyperbola fit is commonly used to calculate one-site binding constants (KD) for the interaction of two proteins or for a protein-ligand interaction. The binding or affinity constant (KD), which is expressed in molarity, can be derived from a hyperbola fit curve by calculating 50% of the response of the y-axis, that is when the first protein is half saturated with the second protein or ligand. In MARS after performing a hyperbola fit, the KD value is equivalent to the ‘b’ value found under the Standard Curve Fit Results.

The figure uses the hyperbola fit and it was derived from data using the SoPRanoTM label-free assay from Pharma Diagnostics. It measure the interaction of varying concentrations of an antibody with an antigen.

Monday, September 17, 2012

FAQ: What is a label-free assay?

Simply put, a label-free assay measures a protein-protein or a ligand-protein interaction without the use of labels or tags on the protein or ligand. There are many applications that measure protein-protein interactions, but they usually require a fluorescent, luminescent, radioactive, or some other kind of tag to be labeled on the protein(s). As one can imagine, those tags could alter the protein-protein binding mechanism, creating a system that is not representative of the native proteins. As an alternative, several label-free methods have arisen that allow the interaction of untagged proteins to be measured.

One of the best known examples of label free technology is BIAcore, which uses surface plasmon resonance (SPR) to measure protein-protein binding. Proteins are conjugated to a microchip, while the interacting protein or ligand is ‘flowed’ over the conjugated protein. Measuring a change in the SPR signal gives detailed information about this interaction, allowing binding constants and rates to be determined. A major drawback to BIAcore and other label-free technologies, though, is the requirement to use expensive, single-use instrumentation.

Recently a new label-free technique has been developed that utilizes a more common piece of laboratory equipment, a simple absorbance spectrophotometer. The SoPRano™ assay Gold Nano Rods from Pharma Diagnostics allow label-free assays to be performed in a microplate-based environment using an instrument that can capture a full UV/Vis absorbance spectrum. Taking advantage of the localized surface plasmon resonance (LSPR) found in Gold Nano Rods which manifests as a spectral curve, the SoPRano™ assay measures protein-protein interactions by monitoring the shift in the wavelength maximum, which is proportional to the size of the interacting protein.

For more information on how to perform the SoPRano™ Label-free assay on BMG LABTECH’s spectrometer-based microplate readers (PHERAstar FS, SPECTROstar Nano, FLUOstar Omega, POLARstar Omega, and SPECTROstar Omega), see the following application notes:

Friday, September 14, 2012

Focus on : MipTec2012


New Technologies that Enhance Cell-based and Label-Free Microplate Assays Will Be Highlighted at Miptec 2012

BMG LABTECH is proud to be a Silver Sponsor of the annual Miptec meeting in Basel, Switzerland from September 24th-27th 2012. The Microplate Reader Company will showcase its newest technology at Miptec 2012, detailing how this technology greatly enhances applications. Stop by Booth #A22 to see unique and proprietary microplate reader technology that separates BMG LABTECH instruments from all of the rest.

As will be highlighted in several posters and at an Industry Symposium, BMG LABTECH will detail how its microplate reader technologies can improve your scientific research. When designing its newest microplate readers, BMG LABTECH analyzed the market to determine what features are missing in a microplate reader that will enhance life science applications. Two such advancements that greatly improve assays are ultra-fast, full spectrum analysis for absorbance assays and Direct Optic Bottom Reading for cell-based assays.

In the Industry Symposium on Tuesday afternoon September 25th, in conjunction with Labcyte and CISBIO, BMG LABTECH will show how its Direct Optic Bottom Reading can create an automated cell-based HTS screening platform that compliments or rivals slower HCS platforms. See how these three companies have brought together their technologies to create such a platform. With Direct Optic Bottom Reading on the PHERAstar FS, the signal-to-blank ratio in fluorescent cell-based assays like GFP, mCherry, and mOrange is more than 300% higher than on fiber optic plate readers. With this level of sensitivity, obtaining higher throughput from an existing FACS or a confocal microscope assay is now possible with this platform.

In addition, BMG LABTECH will highlight in a poster a new label-free system that can determine protein-protein binding kinetics using ultra-fast, full spectrum analysis in a spectrometer-based microplate reader. In collaboration with PharmaDiagnostics, a label-free system that uses their SoPRanoTM label-free technology and a BMG LABTECH spectrometer-based microplate reader has been designed. Without the need to purchase expensive and dedicated equipment, this label-free system can quickly analyze and determine the KD values, EC/IC50s, as well as the association (ka) and dissociation (kd) rates for a protein-protein interaction.

Please visit BMG LABTECH at Booth #A22 at Miptec 2012 to learn more about the future of microplate reader technology today.

For more information and to register for free, visit: http://www.bmglabtech.com/labauto

Thursday, September 13, 2012

Inside Applications: HTS Instrument Discovers Low Affinity Inhibitors of the Inositol Phosphate (IP) Signaling Pathway

IP One Assay Pronciple Point Several low affinity drug 'hits' were only found by the PMT-based reader and not the CCD-based reader
Point PHERAstar FS performs more than twice as fast as the CCD-based reader
Point PHERAstar FS shows superior assay quality parameters such as Z', DeltaF% and assay window
The IP-One HTRF® assay from Cisbio measures the lower response second messenger inositol 1,4,5-triphosphate (IP3). This assay was performed on two different HTS microplate readers with different detection technologies. The data shows that low affinity compounds which were not discovered with a leading HTS CCD camera based imaging microplate reader were readily resolved with the PMT based PHERAstar FS microplate reader from BMG LABTECH.

Wednesday, September 12, 2012

Fun Fact: Happy Birthday Carl Zeiss!

Born 196 years ago yesterday was Carl Zeiss, a German manufacturer of fine optical instruments that are stilled used today. He began manufacturing optical instruments after being trained in medicine and the demand for such precise optical equipment only increased in time. He founded his optical factory in Jena Germany in 1846 and his reputation for making high quality optics became known. He died 72 years later in 1888. Find out more about Carl Zeiss here: http://en.wikipedia.org/wiki/Carl_Zeiss

Tuesday, September 11, 2012

Did you know that MARS has Confidence Intervals on its standard curve tab?

Yes. Using a drop down menu in MARS Data Analysis Software, options such as showing replicates, error bars, and now Confidence Intervals (95%) can be calculated. To indicate the intervals, two dotted lines are on the standard curve graph. To see the min and max values, hit the Fit Parameters to call up the values in a separate window. This is one way BMG LABTECH continually strives to update its software.

To find out more about BMG LABTECH Software, visit http://www.bmglabtech.com/products/software.cfm

Monday, September 10, 2012

FAQ: What is the difference between a Life Science microplate reader and a High-Throughput Screening (HTS) microplate reader?


Basically sample throughput HTS instruments can measure 96 or lower microplates, but they are designed to measure higher density microplates like 384-, 1536- and 3456-well. Life science instruments, on the other hand, are designed to measure 6- to 96- well microplates. Most life science instruments can still measure 384- or 1536-well microplates, but they are designed for lower density formats.  HTS instruments are also designed to measure microplates faster than life science readers, in order to increase throughput.

Most assays and applications can be measured on both categories of instruments, but HTS microplate readers usually have a lower limit of detection, therefore lower concentrations can be used.

For more information on the different BMG LABTECH microplate readers visit: http://www.bmglabtech.com/products/microplate-reader.cfm

Friday, September 7, 2012

Focus On: Microplate-based Fluorescence and Luminescence Assays for Calcium Flux Measurements.

Measuring the change in calcium flux in cell-based assays is a very useful tool for monitoring the downstream effects of receptor activation or inhibition. For instance, activated Gαq coupled G-protein coupled receptors (GPCRs) such as the Serotonin 5-HT2, Histamine H1, or the Vasopressin 1 receptors, will subsequently activate the phosopholipase C (PLC) signaling cascade, which ultimately, amongst other things, results in an intracellular calcium flux.
As a way to study the activation of such receptors connected to changes in calcium flux, several different luminescence and fluorescence assays have been developed that can work in a microplate-based environment. Calcium flux assays work the best on microplate readers that can measure and inject  into a well at the same time. Since calcium flux assays occur extremely fast, instruments only have seconds to capture the necessary measurements. With simultaneous injection and measurement, every data point is captured before, during, and after injection.

Over the years, many different calcium signaling compounds have been created that can be used on microplate readers. Here is a brief and unfinished list of some of the more popular calcium flux assays on the market:

- Fura-2, Fluo-3 were first generation dyes
- Fluo-4 Direct™
- Fluo-8
- Aequorin Luminescence
- Clonetics™ luminescence primary sensor
- FLIPR® fluorescence calcium assay kit
- Oregon® Green

The following BMG LABTECH microplate readers can perform calcium flux assays, LUMIstar, FLUOstar and POLARstar OPTIMA; LUMIstar, FLUOstar and POLARstar Omega; PHERAstar FS; and the NOVOstar.

Thursday, September 6, 2012

Thursday Application – A Calcium Bioluminescence Assay: A New Research Based Application Video


A research based application video entitled, A Calcium Bioluminescence Assay for Functional Analysis of Mosquito (Aedes aegypti) and Tick (Rhipicephalus microplus) G Protein-coupled Receptors, has been published on JoVE, the Journal of Visualized Experiments, and it uses the NOVOstar microplate reader. This video publication shows in detail how researchers at Texas A&M produced a stable cell line in mammalian CHO cells to investigate GPCRs from two different anthropods, the southern cattle tick Boophilus microplus (Canestrini), and the mosquito Aedes aegypti (Linnaeus). The NOVOstar microplate reader was used to study the GPCR activated calcium response of these stably transfected cell lines using a bioluminescent signal. As the authors note:

“This method has three primary applications. First, the technique can be applied for receptor deorphanization through ligand activity measurements. Second, the assay can resolve ligand-receptor structure-activity relationships (SAR). Third, the methods can be used in drug discovery. Furthermore, this protocol can be used to study the activity of agonists or antagonists on almost any GPCR.”

View all BMG LABTECH videos here and view the research based application videos here.

Tuesday, September 4, 2012

Did you know that BMG LABTECH will be represented at numerous conferences in the coming weeks?


The breadth of topics shows the versatility of BMG LABTECH multimode microplate readers.

If you are in the area, stop by one these venues in September to see BMG LABTECH and the future of microplate reader technology, today:

4-5th - Genomics Research in Frankfurt, Germany
5-6th - ELRIG in Manchester, UK
6-9th - Free Radical Research International in London, UK
9-12th - European Calcium Society, Toulouse, France
10-13th - Black Forest Retreat on Molecular Plant Science, Black Forest, Herzogenhorn, Germany
12-14th - British Association for Cancer Research, Harrogate, UK
17-19th - RSC/SCI Symposium on GPCRs in Medicinal Chemistry, Windlesham, UK
19-21st - HTS Thematice school in French: Ecole Thematique de criblage, Toulouse France
23-27th - ComBio, Adelaide, Australia

See all BMG LABTECH upcoming events here.

Please contact your local representative they may be able to help with entrance to any of the above shows.

Monday, September 3, 2012

FAQ: Why do you need different microplates when measuring DNA and protein using 260 and 280 nm absorbance measurements, respectively?


When performing absorbance measurements in the UV range (<400 nm), UV clear microplates are needed for accurate measurements. The most common microplates are made of polypropylene or polystyrene and these materials absorb light in the UV range. Therefore, DNA/protein quantitation that require measurements at 260/280 nm need a material that will not absorb UV light.

There are several types of UV-transparent microplates, including: