Thursday, January 31, 2013

Applications Thursday: Discovery of Low Affinity Inhibitors of IP signaling using HTRF

G-protein coupled receptors (GPCR's) represent a major class of cellular signaling receptors that play a vital role in a number of physiological responses. As such they are a vital target to be analyzed for drug discovery. The PHERAstar FS from BMG LABTECH is a great tool for this type of analysis as exemplified in the application note: HTS Instrument Discovers Low Affinity Inhibitors of the Inositol Phosphate (IP) Signaling Pathway

Biosynthesis of IP1
One of the second messengers that is produced upon GPCR activation is IP3 which in turn leads to calcium release. However, since IP3 has a very short half life detection of a downstream metabolite IP1 is prefered. This is the basis of  Cisbio's HTRF IP-One assay which employs monoclonal antibodies raised to IP1 which are linked to a donor molecule and IP1 linked to an acceptor molecule. In the absence of endogenously produced IP1 energy transfer will occur between donor and acceptor. Stimulation of IP1 production will lead to competition with the IP1 linked to an acceptor and a resulting decrease in FRET. Using the dual emission detection capabilities of the PHERAstar FS you can monitor not only the emission due to FRET but also emission of the donor when FRET does not occur and determine the ratio of these emissions.

HTRF® ratios obtained for the IP-One assay with the PHERAstar FS
The 1536 well screen exhibits a robust assay window and reveals hits that are 20-30% of maximal inhibition that represent compounds with excellent potential as drug candidates.

For more information please visit BMG LABTECH where you can read this application note and many others.

Wednesday, January 30, 2013

Fun fact: In the past 2 years 491 new species have been described

Scientists from the 10 Senckenberg Gesellschaft für Naturforschung institutes consider taxonomy and scientific collections among their most important focal points. The objective being to track biodiversity and see that it is maintained.

 Variation in coloration pattern in life of Diasporus citrinobapheus from different localities:
http://www.pensoft.net/J_FILES/1/articles/2774/2774-G-3-layout.pdf
Since more than 100 species die out each day it is important to identify new species so that they can be protected. Although only recently discovered, many of these species can already be considered endangered.



Senckenberg researchers discovered 87 fossilized species but the majority are living. Most of the new species are arthropods but molluscs and plants are also highly represented. In addition, the majority are terrestrial species and the continental distribution has Asia leading the way with 324 newly discovered species.

For more information on this report visit this science daily article:

Almost 500 New Species Discovered at Senckenberg: Newly Discovered Species in 2011 and 2012

Tuesday, January 29, 2013

FAQ: What do the experts say about the PHERAstar FS

To find out you can visit select science!

The self proclaimed: '...fastest way to impartial, expert opinion about the best laboratory equipment...' is featuring the PHERAstart FS from BMG LABTECH in their product reviews section today. There you can watch a video or read about the characteristics of the PHERAstar FS that make it the gold standard HTS microplate reader and read reviews from expert users.

The select science description of the PHERAstar FS  includes some of this microplate readers important features such as:

Tandem spectrometer/filter based technology
Two lasers for assay selective excitation
Ability to perform precise kinetics
Top and bottom reading capabilities
Cell-layer scanning
Access to MARS data analysis

And more!

In addition there are links to application notes that will begin to give you an idea about the versatility of the PHERAstar FS. For more information you should visit the BMG website and explore all the capabilities of the PHERAstar FS and the applications to which it can be employed.


Monday, January 28, 2013

Did you know BMG LABTECH gave away an iPad mini at SLAS?

BMG LABTECH in cooperation with other vendors at SLAS 2013 in Orlando gave attendees the opportunity to enter to win a number of great prizes. As our contribution BMG provided an iPad mini!

Apple iPad mini in black, image rendered
http://www.pixeden.com/psd-mock-up-templates/ipad-mini-psd-vector-mockup
The winners, chosen from over 80 participants were:


Michelle Arata, Ra Pharmaceuticals (iPad mini from BMG LABTECH)
Yan Zhai, Eli Lily (iPad mini from Cell Dynamics)
Lily Shi, Genentech (MacBook Air from Promega)

Congratulations to all winners and thank you to everyone that participated!

Friday, January 25, 2013

Focus on: PHERAstar based primary screens at Sanford-Burnham

The PHERAstar FS  from BMG LABTECH is a highly versatile instrument. This is evident from the variety of applications that the Burnham Center for Chemical Genomics has employed in the design of primary screens using the PHERAstar FS. They have studied post-translational modification, mitochondrial permeability, protease activity and protein-protein interactions and have performed these screens using TR-FRET, fluorescence intensity, absorbancefluorescence polarizationHTRF or FRET detection all on one microplate reader!
The PHERAstar FS from BMG LABTECH

For more information on the primary screening assays please use the following links to the appropriate pages at PubChem:

uHTS identification of inhibitors of cullin neddylation in a TR-FRET assay [Primary Screening]

Single concentration confirmation of uHTS inhibitor hits of the mitochondrial permeability transition pore via a fluorescent based assay [Primary Screening]

Single concentration confirmation of uHTS inhibitor hits of Sentrin-Specific Protease 8 using Nedd8 Protein Substrate [Primary Screening]

uHTS identification of small molecule inhibitors of the mitochondrial permeability transition pore via an absorbance assay [Primary Screening]

uHTS identification of modulators of interaction between CendR and NRP-1 using Fluorescence Polarization assay [Primary Screening]

Single concentration validation of uHTS RPN11 inhibitor hits using a MMP-2 Fluorescence assay [Primary Screening]

Single concentration confirmation of uHTS inhibitor hits from RPN11 in a Fluorescence Polarization assay [Primary Screening]

uHTS HTRF interference counterscreen assay for validation of inhibitors of SUMOylation [Primary Screening]

Identification of compounds inhibiting the binding between the RUNX1 Runt domain and CBFb via a time resolved fluorescence resonance energy transfer (TR-FRET) assay. [Primary Screening]

uHTS identification of compounds inhibiting the binding between the RUNX1 Runt domain and CBFb via a fluorescence resonance energy transfer (FRET) assay. [Primary Screening]

Single concentration confirmation of uHTS for the identification of UBC13 Polyubiquitin Inhibitors via a TR-FRET Assay [Primary Screening]

uHTS identification of UBC13 Polyubiquitin Inhibitors via a TR-FRET Assay [Primary Screening]


Thursday, January 24, 2013

Applications Thursday: A Novel Fluorescent Intensity uHTS Assay that Utilizes Physiological Protein Substrates

A recent assay performed at Sanford-Burnham Center for Chemical Genomics entitled: 'Single concentration confirmation of uHTS inhibitor hits of Sentrin-Specific Protease 8 using Nedd8 Protein Substrate' describes a fluoresence intensity assay using BMG LABTECH's PHERAstar FS designed to find inhibitors of an enzyme involved in the neddylation pathway. Targeting the neddylation machinery is attractive due to the role of this machinery in indirect regulation of cell cycle progression, DNA replication and DNA damage response mechanisms.

DNA_damaged_by_carcinogenic_2-aminofluorene_AF_.jpg
The application is straightforward enough involving detection of deconjugated aminocoumarin from a Nedd8-AMC substrate. The released aminocoumarin is readily detected with ex/em at 350/450  using a microplate reader such as the PHERAstar FS from BMG LABTECH. Using the full length Nedd8 should reveal inhibitors that bind to the actual conformation of SENP8 when it is bound to substrate.


Wednesday, January 23, 2013

Fun fact: Growing willows diagonally yields more sugar for biofuel production

A collaboration between the Imperial College of London, Rothamsted Research and Orkney College have discovered a genetic trait in willows that yields the production of more glucose when the trees are grown at an angle. The production is normally a response to growth in windy conditions which would strengthen the trunks of the trees.

The authors believe that they can now breed trees that contain the appropriate genetic characteristics to yield plants with higher capacity in biofuel production.

Tuesday, January 22, 2013

Did you know over 60 screening procedures that use BMG instruments are freely available?

Pub chem is a collection of freely searchable databases designed to provide information about small molecules and their biological activity. The databases are: Compound, Substance and BioAssay.



Using the PHERAstar  series of microplate readers from BMG LABTECH the Sanford-Burnham Center for Chemical Genomics has generated 68 entries and counting!


Monday, January 21, 2013

Happy MLK Day!

Many workers around the United States are allowed the day off from work in the national observance of the birth of Martin Luther King Jr. His actual date of birth is January 15.

 In observance of his birthday BMG LABTECH suggests that you provide some service to your community to reflect the dedication MLK had to his community during  his life.

Martin Luther King, Jr. showing his medallion received from Mayor Wagner1964. Library of Congress, New York World-Telegram & Sun Collection, Reproduction Number LC-USZ62-126558. Phil Stanziola, NYWT&S staff photographer

Friday, January 18, 2013

Focus on: SLAS wrap up

It was great to see so many of you attending SLAS this week in Orlando, FL. There was plenty of excellent science and technology on display and BMG was happy that we were able to participate as a diamond sponsor for the event. Attendees came from around the world to take part in the 4 day event.

We at BMG were especially excited to meet with Hayley Jones and David Whalley, two scientist at MRC Technology in London. Both have been doing some exciting work with our PHERAstar FS and took the opportunity to share their work at SLAS.

Illustration of the pain pathway in René Descartes' Traite de l'homme (Treatise of Man) 1664. The long fiber running from the foot to the cavity in the head is pulled by the heat and releases a fluid that makes the muscles contract.
Hayley gave a poster entitled: Identification of and Characterisation of Novel Positive Allosteric Modulators of the Galanin 2 Receptor. The galanin 2 receptor (GalR2) is implicated in regulating pain reception, especially in regard to chronic nerve damage. However, since there are no high affinity small molecule compounds that activate GalR2) Hayley is investigating potential PAM's (positive allosteric modulators). Employing the HTRF IP-One assay from Cisbio and the PHERAstar FS from BMG LABTECH Hayley has succesfully performed the intitial screeen of 100,000 compounds and is now looking to perform some dose response analysis on the compounds. It is hoped that identification of PAM's for GalR2 will have therapeutic advantages including the retention of physiological response.

  • purine (spacefill)
David is in the early stages of his project and gave a poster entitled: Development of a High-Throughput Screening Assay to Identify Inhibitors of the de novo Purine Biosynthetic Pathway. He is looking for inhibitors of a bi-functional enzyme that is overexpressed in breast cancer. Since this enzyme is ATP dependent he was able to employ the Transcreener assay from Bellbrook Labs with detection performed by the PHERAstar FS to perform a high-throughput screen. He is now ready to screen a compound library with 100 K to 200 K compounds.

Great job by everyone in attendance! See you next year in San Diego!



Thursday, January 17, 2013

Applications Thursday

The technologies of BMG LABTECH, DiscoveRx and Labcyte have combined to create an approach that will allow you to screen for activation of G-protein coupled receptors or tyrosine kinase receptors. The application employs a sensitive assay, sub-microliter volume liquid handling, and the capabilities of the PHERAstar FS microplate reader.

 PathHunter® assay principle

The assay technology from DiscoveRx is an adaptation of enzyme fragment complimentation that provides a novel cell-based assay format for detecting protein-protein interaction. Activation of the receptor upon ligand binding allows interaction of GPCR-β-Arrestin or RTK-SH2 protein resulting in a measurable chemiluminescent signal.

The application note: Nano High-Throughput Screening (nHTS) Platform-Miniaturization of Cell-based GPCR and Kinase Assays describes proof of principal that will allow you to maximize your results by using fewer cells and smaller reagent volumes.


Wednesday, January 16, 2013

Fun fact: Precise altering of genomes may now be possible

Two recent articles both published in Science describe an approach that could be used to edit genomes. One is  entitled: 'Multiplex Genome Engineering Using CRISPR/Cas Systems' the other: RNA-Guided Human Genome Engineering via Cas9This approach could be useful for applications such as designing organisms for bio-fuel production.

The approach takes advantage of naturally occurring bacterial protein-RNA systems that use the RNA component to recognize viral DNA sequences and the enzyme Cas9 to cut the DNA. By adding new RNA sequences; specific DNA sequences can now be targeted for cutting. The result is a tool whose application could extend to new therapies for diseases such as Huntingtons disease, which appears to be caused by a single abnormal gene.
Single striatal neuron (yellow) transfected with nuclear inclusion (orange) mHtt, other neurons in background (blue), from press release [1], Attribution 'Dr. Steven Finkbeiner, Gladstone Institute of Neurological Disease, The Taube-Koret Center for Huntington's Disease Research, and the University of California San Francisco'

Tuesday, January 15, 2013

Did you know cell based screens can be performed on as few as 250 cells/well?

It is now feasible to perform cell-based assays in sub micoliter volumes using as few as 250 cells due to improvements in assay sensitivity, liquid handling and microplate reader technology. The PHERAstar FS from BMG LABTECH, The Microplate Reader Company, was used in conjunction with technologies from DiscoveRx, and Labcyte to show that analysis of G-protein coupled receptor function could be performed in 3456 well plates. Specifically, suspension CHO cells at different densities, which express the dopamine receptor, were tested for their ability to respond to agonist. Results indicate that 250 cells in less than 1 microliter exhibit a clear response.

3456-well Protocol Optimization. PathHunter® DRD2L cells 100 or 250 cells/well (in suspension) were added to the AURORA 3456 well plates in 0.5 or 1 μL volumes via the Echo® instrument. Compound (5nl of 100X in DMSO) was added to the cells plated in 0.5 µl (10 nl for the cells plated in 1µl) via the Echo® instrument.

With high through put assays of this type it will be possible to perform more screens with lower reagent costs.

Monday, January 14, 2013

FAQ: Can cell based assays be performed using 3456 well microplates?

By combining the technologies from DiscoveRx, Labcyte and BMG LABTECH we can now answer: YES!

The application note: Nano High-Throughput Screening (nHTS) Platform -Miniaturization of Cell-based GPCR and Kinase Assays describes the feasibility of performing cell-based assay in sub microliter volumes that can be detected with the PHERAstar FS. This results in decreased amount of test compound used and thus means the experiments are economical with regard to both money and time. The results exhibit good correlation with results from other well formats and displayed reproducibility.

Reproducibility of 3456-well miniaturization of the PathHunter® β-Arrestin CHO Cells
 Plots A and B are two different measurements taken on different days

Friday, January 11, 2013

Focus on Events: PAG 2013

You can visit BMG LABTECH, the microplate company, at booth 613 during PAG XXI from January 12-16 in San Diego, CA. There you will see the versatility of BMG instrumentation on display and be able to interact with our representatives.

BMG will also be presenting data that shows how the PHERAstar can be used in high speed, FRET based SNP genotyping. SNP's have become an invaluable tool for genetic research and are employed in a wide range of scientific fields. BMG's optimized optical modules result in improved speed and accuracy of your SNP analysis.


Thursday, January 10, 2013

Did you know that non-binding plates should not be used in the NEPHELOstar Plus because it increases light-scattering and thus the blank?



The NEPHELOstar Plus uses a high-intensity light source which is directed through a sample. Particles in the sample are detected based on the amount of light scatter that occurs. This detection is approximately 30 times more sensitive than readers that detect a reduction in direct light. The increased sensitivity makes the NEPHELOstar Plus useful for a number of applications including: drug solubility screening and growth curve monitoring for microbes.

 However, the high sensitivity can make this instrument susceptible to detecting high background when anything, besides the sample, that can scatter light is also present. This includes the substances and other coatings that are used to make non-binding microplates effective. In our tests, non-binding plates increased the blank 90 times that seen with non-coated plates. To obtain great results using the NEPHELOstar Plus stick with untreated plates of up to 384 wells for your assays.

Wednesday, January 9, 2013

Fun fact: After 50 years; mode of action of highly prescribed diabetes drug may now be understood

The most frequently prescribed drug for treatment of type 2 diabetes, metformin, belongs to a class of drugs called biguanides. Despite their effectiveness, the mechanism of action for these drugs has been poorly understood. That changed with the publication in Nature of: Biguanides suppress hepatic glucagon signalling by decreasing production of cyclic AMP. The paper was produced by the lab of Morris J. Birnbaum at the University of Penn with contributions from authors in France.

It had been suggested that metformin results in reduced glucose synthesis by activating AMPK but recent evidence showed that mice lacking AMPK still responded to metformin treatment.This led to the current findings that metformin acts by increasing the levels of cellular AMP which has the effect of inhibiting andenylate cyclase and therefore decreasing cAMP production. The discovery of this novel mechanism may lead to new treatments targeting adenylate cyclase.

Adenylate cyclase by en:user:Bensaccount
  • 23:54, 26 April 2006 Bensaccount 452x258 (60841 bytes) (Adenylate cyclase by [[:en:user:Bensaccount]])


Tuesday, January 8, 2013

Did you know that BMG LABTECH microplate readers have an advanced script mode that can be used to combine multiple functions of the plate reader easily?

Using script mode, researchers have full control of how their protocol is used to measure samples. This function is in addition to the easy to manage Testrun Protocol setup and is especially useful when creating intricate kinetic measurements combining shaking and resting periods, multiple measurement modes, and more. For example, the new assay for prion measurement, Real-Time Quaking Induced Conversion Assay (RT-QuIC), takes advantage of the script mode by combining a temperature setting of 42°C with alternating shaking and resting periods of one minute with measurements every 15 minutes. Advanced measurement protocols such as this can be easily created for use on BMG LABTECH readers.


Script Wizard allows BMG LAGTECH users to multiplex protocols for complex assays like RT-QulC prion seeding assay.

Monday, January 7, 2013

FAQ:How flexible are the microplate readers from BMG LABTECH?


BMG LABTECH microplate readers have unlimited flexibility with temperature control to 65°C and shaking with variable speeds in three directions - orbital, double orbital, and linear. Readers can take measurements from the top or bottom of a plate. Measurements can be endpoint or kinetic. Kinetic can be expanded to plate mode, which reads all designated wells several times over a longer period of time, or in well mode, which reads a single well several times quickly and then moves on to the next well. In addition, readers can include basic modes such as fluorescence , absorbance, and luminescence, or more advanced modes such as time-resolved fluorescence, TR-FRET, and AlphaScreen®. Our advanced control software easily allows users to create testrun protocols combining set temperature, shaking, and reading modes. But if you need more than the basics, the script mode can easily control the most advanced of protocols. One script used in measuring prion activity, for example, easily combines repetitive shaking of 700 rpm in a double orbital orientation for a minute, and a minute of rest, as well as a measurement every 15 minutes for several days.

 
BMG LABTECH instruments can shake the microplate in three different way - linear, orbital, and double orbital

Friday, January 4, 2013

Focus on Events: SLAS 2013

Visit BMG Labtech, the Microplate Reader Company, at booth 721 at SLAS 2013, January 12-16, in Orlando, FL. For this 2nd annual conference BMG is proud to be a Diamond Sponsor and we will be participating in a number of events that will show how we are engineering a better biological solution.

The versatility of our line of microplate readers will be on display and a number of tutorials, posters and collaborators booths will feature the PHERAstar FS.

On Monday, January 14 at 12:30 BMG will co-present at a tutorial sponsored by Labcyte that will discuss the miniaturization and automation of Cisbio's HTRF assays. Then on Tuesday, January 15 at 12:30 BMG will host a tutorial that will show how our microplate technology can enhance  assay development and high through-put screening. In addition, posters that highlight the performance of the PHERAstar FS and NEPHELOstar in specific applications will also be presented.

Wednesday, January 2, 2013

Fun fact: Your New Years resolution could do more harm than good

As we begin the new year many people have made a resolution to lose weight with a diet and exercise. While this is a good choice, as with many things you should proceed caution as described in the article: Five Reasons Why New Year's Resolutions to Diet and Exercise Might Be Unhealthy 

Make sure that you are aware of the potential allergens that are present in some of the prepared meals offered by diet plans which may hinder your new plan. Furthermore you should be aware that allergens are not only present outdoors, and that the surfaces of the workout equipment or even your clothing could lead to problems. This is not to say that exercise should be avoided. But you should be aware of potential pitfalls so that you can have the best experience and get the most out of your resolution.


By SariSabban [CC-BY-SA-3.0 (http://creativecommons.org/licenses/by-sa/3.0)], via Wikimedia Commons
Simplified diagram showing key events that leads to allergy initiation

Tuesday, January 1, 2013

Happy New Year!

BMG LABTECH wishes everyone a Happy New Year! 

After a great year in science in 2012, we can only imagine what will come in 2013! Here is a great article in Nature describing some of the fantastic scientific happenings of 2012. From the Higgs Boson discovery to the Mars Rover landing to the restructuring of pharmaceuticals, it was a banner year for science.

File:CMS Higgs-event.jpg
Higgs-Boson Event Simulation by Lucas Taylor.