Use of Far-Red Emitting DNA Dye DRAQ5 for Cell Cycle Analysis with Microplate Cytometry
Poster Sep 14, 2007
Sarah Payne, Paul Wylie, Roy Edward and Andrew Goulter
There is an increasing demand for multiplexing in high content assays to maximise data generation and allow correlation across multiple readouts.
Currently, the degree of multiplexing can be limited by the available reagents, with the majority of fluorescent probes optimised for excitation at 488 nm. The use of fluorescent probes with spectral profiles that overlap hinders their use in multiplexing assays. Another limitation is that many fluorescence detection systems excite and detect over a narrow wavelength range, making them incompatible with certain probes.
Laser-scanning fluorescence microplate cytometers, such as the Acumen® eX3 (TTP LabTech Ltd, Melbourn, UK), offer 405nm, 488nm and 633nm laser excitation in a single instrument. This technology is heavily used in oncology research including cell proliferation and cell cycle analysis using the DNA stains propidium iodide (488nm excitation) and Hoechst 34580 (405nm). Here, we describe the use of DRAQ5™ (633nm) on an Acumen eX3.
Use of DRAQ5 has become popular since it is a far-red fluorescent DNA dye that can be used in live and fixed cells in combination with other common fluorophores, especially GFP fusions and FITC-tags without spectral emission overlap. Thus DRAQ5 offers great potential for multiplexing DNA content analysis with immunodetection assays.
Knockout of microRNAs Using the CRISPR-Cas9 System with Paired Synthetic crRNAsPoster
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE
Exploiting Polypharmacology in Precision Oncology: Identification of Differential Kinase Off-targets Among Clinical PARP InhibitorsPoster
Can we use computational methods to identify previously unknown off-targets of PARP inhibitors that can explain their observed differences?READ MORE
Bioluminescent Assay for GTPases Allows Measurement of GTPase, GAP and GEF ActivitiesPoster
We have developed a homogenous bioluminescent assay (GTPase-Glo) system to analyze these proteins in a simple, convenient “add-mix-read” format.READ MORE
5th edition of the International Conference Clinical Oncology and Molecular Diagnostics
Jun 17 - Jun 18, 2019