|Label free technology: the weird and wonderful world of cell signaling|
Ryall A†, Shearer J†, Brodbeck D*, Nufer O*, Cronk D† and Fasler S*
Whilst conducting an evaluation of label free technology platforms we
encountered a number of surprising, but reproducible, observations. For
example, receptor over-expression impacts endogenous signaling
responses; surface coatings affecting cell adherence can change the
response profile; and it is important to choose the appropriate analysis
metric for complex profiles. Awareness of the subtleties of cell signaling
within model systems such as CHO and HEK recombinant over-expression
|Label free technology: value added information in the drug discovery process|
Brodbeck D*, Nufer O*, Ryall A†, Shearer J†, Fasler S* and Cronk D†
We have evaluated Molecular Devices CellKey™ and SRU BIND® label free
detection technologies and their application to hit finding and hit validation
using cell based assays. The presented data highlights:
|Analysis of migration using the OrisTM Cell Migration Assay-TriCoated kit on the POLARstar Omega.|
The Oris™ Cell Migration Assays from Platypus offer certain advantages over traditional migration assays. Especially the TriCoated kits enable the researcher to test the effect of different extracellular matrix components like Collagen I or Fibronectin on migration. In this application note we show the real-time monitoring of cell migration using a POLARstar Omega from BMG LABTECH equipped with a CO2 gas vented system.
|The Transcreener® ADP2 Universal Kinase Assay from BellBrook Labs is readily performed on BMG LABTECH microplate readers using different assay formats|
The Transcreener® ADP2 FI assay kit from BellBrook Labs is a simple one-step competitive red fluorescence immunoassay based on the detection of ADP. In this application note we show that this assay is compatible with four different microplate readers from BMG LABTECH. With the PHERAstar FS and Plus, as well as the POLARstar and FLUOstar Omegas comparable standard curves and EC50 values were obtained.
|A novel method for coupling capillary columns|
Wil van Egmond, Daniela Peroni,
Using a proprietary, low-melting, deactivated glass we have developed a novel instant Meltfit™ connection. Reliable connectors are made in-situ in less than a minute.
|A Comparison of AlphaLISA Bead-Based Luminescence and Electrochemiluminescence Immunoassay Technologies for Detection of Human EPO, Amyloid Beta 42 and VEGF in Complex Sample Matrices|
Anuradha Prasad, Ashleigh Price, Stephen Hurt, Rathnam Chaguturu
The University of Kansas in collaboration with PerkinElmer Inc. worked on looking at the comparison of the AlphaLISA Technology and an Electrochemiluminescence Technology to measure assay windows, lower and upper detection limits and intra - and inter-assay precision. In this study, three AlphaLISA no-wash assays, which employ a faster and less complex protocol, were found to deliver highly sensitive and accurate results, equivalent to those obtained in ECL technology.
|Real-Time Monitoring of ATP Depletion Suitable for HTS|
ATP depletion assays of kinases, ATPases and aminoacyl tRNA synthetases can be performed in the presence of firefly luciferase/luciferin, by monitoring the light emission continuously. Such assays are preferably set up to result in a first order reaction rate with respect to ATP consumption.
|Microcarrier based production of cryopreserved cells|
Sharon Davies, Lynne Smith, Sian Kalinka, Philip Meyler, Chris Jones, Rahman Ismail, Elizabeth Price, Stephen Game, J M Kendall., GE Healthcare
The use of cellular assays in drug screening continues to grow with over 50% of primary screens using cell based formats in 2006. The majority of cells used in the drug discovery industry are fresh, using ‘just in time’ batch processing from in house facilities. This process could give rise to a number of issues, namely batch variation, scheduling of cell production and capacity management. We describe the use of microcarrier technology in combination with a suitably configured bioreactor to pro
|Smart Solutions in drug discovery|
Ian Kalinka, Lynne Smith, Elizabeth Price, Angela Marenghi, J.M.Kendall, Ray Ismai
Here we present data on the development of a large scale production process for the provision of two different cryopreserved transiently transfected cell lines. Included is identification of a suitable transfection reagent that is efficient, non-toxic and cost effective. This process has been validated using these two cell lines and two different cell-based assays.