Latest Application Notes
A Leukocyte Adhesion Assay Performed on BMG LABTECH’s FLUOstar OPTIMA
Gene regulatory networks were used to identify novel regulatory hubs genes involved in the inflammatory response in human umbilical endothelial cells (HUVECs). To assess the potential role of these genes a leukocyte adhesion assay was utilized to screen for potential inflammatory markers. The FLUOstar OPTIMA microplate reader from BMG LABTECH was used for cell based bottom optic measurements and well scanning. The results show good reproducibility and can be adapted to e.g. primary neutrophils.
Tag-liteTM: New HTRF® cellular platform for cell surface receptors’ study and screening
In a recent publication, Maurel et al* presented the combination of Cisbio’s HTRF and Covalys’ SNAP-and CLIP- Tag technologies to investigate cell-surface protein-protein interaction. In this study, an application for GPCR oligomerization was accurately documented and the advantages of this new approach presented.
Size-selective assessment of tight junction paracellular permeability using fluorescently labelled dextrans
A method for the quick and reliable analysis of size-selective paracellular tracer diffusion using fluorescent dextrans of different molecular weights is described. The assay could be easily adapted to large scale screens to identify genome-wide regulatory pathways or small molecules to modify junctional permeability. Using BMG LABTECH´s multimode plate reader FLUOstar OPTIMA for these experiments allows for quick and consistent fluorescence measurements in either kinetic or endpoint format.
Detection of NADH and NADPH with the Omega’s High Speed, Full UV/Vis Absorbance Spectrometer
NAD+, NADH as well as NADP+, NADPH have been known to play vital roles in energy metabolism, antioxidation, and reductive biosynthesis. Changes in these systems can be monitored using absorbance spectroscopy. NADH and NADPH absorb light at 340 nm, whereas NAD+ and NADP+ do not. The FLUOstar, POLARstar and SPECTROstar Omega microplate readers from BMG LABTECH all have a UV/Vis spectrometer that can measure any absorbance range from 220-850 nm.
Measuring Mitochondrial Membrane Potential using the FLUOstar Omega Microplate Reader
Measurement of the mitochondrial membrane potential is useful in a variety of research areas. Mitochondrial membrane potential is usually measured using non-invasive cationic dyes. These dyes are sequestered into the mitochondrial matrix in amounts directly proportional to the membrane potential. BMG LABTECH’s FLUOstar Omega was used in an hts assay measuring mitochondrial membrane potential in human cells. The results presented here were gained using primary human fibroblast cells from controls
Evaluation of PARP Inhibitors: Performed on BMG LABTECH’s FLUOstar Omega
Poly (ADP-ribose) polymerases (PARP) transfer ADP-ribose to itself and other nuclear proteins such as histones, therefore PARP play a crucial role in regulating DNA repair. A luminescent cellular PARP assay and a fluorescent cell viability assay were performed on BMG LABTECH’s FLUOstar Omega to evaluate PARP inhibitors. PARP inhibition has been demonstrated to potentiate the cytotoxicity of anti-cancer drugs and ionising radiation. The ability of the FLUOstar Omega to measure absorbance, lumines
Nephelometric Monitoring Growth of Candida albicans Using BMG LABTECH’s NEPHELOstar
Laser nephelometry has been shown to be a reliable technique for the measurement of drug solubility in 96-well plate format. In this note the use of BMG LABTECH´s NEPHELOstar to investigate the effects of complexation on the drug antimycotic activity is described. Laser nephelometry can distinguish between the concentration at which the drug just goes into or just comes out of solution. This is presented in phase solubility studies. Furthermore nephelometry was used to monitor fungal growth.
New Transcreener® ADP2 FP Assay performed on BMG LABTECH’s PHERAstar Plus HTS Microplate Reader
Transcreener™ ADP2 Assay kits are far-red competitive fluorescence polarization immunoassays based on the detection of ADP. Any enzymatic reaction that uses ATP in a range from 0.1-1000 µM can be monitored. With its dual emission detection and five photomultiplier tubes, BMG LABTECH´s PHERAstar Plus provides the speed and sensitivity needed to take full advantage of BellBrook Labs Transcreener™ technology. Furthermore, a specific optical module was developed, thereby making assay setup simple.
Africa and Design of Experiments
Reaction optimisation work using traditional techniques can be complex, time consuming and expensive, but is a vital part of the development of all chemical processes. One approach is the use of statistical experimental design (DoE). This application note gives some guidance on DoE and it’s use with the Africa system.