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Nucleofection® Enables Luciferase Reporter Assays in Primary Cells and Virtually Any Cell Line
Amaxa GMBH
The combination of Promega’s luciferase products and amaxa's Nucleofector® Technology enables researchers to obtain much more significant results with physiologically relevant cells. In this technical note, we give suggestions for Nucleofection® of luciferase constructs and an overview about the wide range of possible applications.
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Role of the small GTPase Rho in the regulation of the cytoarchitecture of dendritic cells
Amaxa GMBH
Dendritic cells (DC) are critically involved in the initiation of innate and adaptive immunity. The wide variety of T cell-mediated responses promoted by DC is a direct consequence of their functional plasticity. An important determinant for DC biology and effector functions is their unique morphology, which contributes to their migratory, antigen-capturing and T cell-stimulating activities. The availability of efficient, reproducible and non-toxic methods for DC transfection would allow the ana
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Nucleofection®: A Method to Stably Transfect C1R Cells With MHC Class I Genes
Amaxa GMBH
Major histocompatibility complex (MHC) class I molecules are heterotrimeric cell surface glycoproteins found on most nucleated cells of the body displaying endogenously synthesized peptides. Cells displaying class I molecules with healthy host-derived peptide ligands remain unharmed while those presenting peptide ligands characteristic of infected or neoplastic cells become targets for the immune response. In humans the main MHC class I subfamilies are the Human Leukocyte Antigen (HLA) loci A, B
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Nucleofection® – High Transfection Performance and Superior Preservation of Functionality
Amaxa GMBH
Nucleofection® has become the method of choice whenever transfection of primary cells or hard-totransfect cell lines is required. In this application note we show that Nucleofection® of frequently used primary cells results in highly efficient transfer of DNA and other substrates while at the same time maintaining excellent cell viability and post transfection functionality.
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Achieving Efficient Delivery of Morpholino Oligos with Nucleofection®
Amaxa GmbH
Morpholino oligos provide a stable, specific and effective antisense activity. Developmental biologists have long since used these properties by microinjecting or electroporating Morpholinos into embryos to allow for splice-blocking or gene knockdown. However, efficient delivery of Morpholinos in mammalian cell cultures can be a challenge. While complex formation with lipid-based reagents is ineffective due to the non-ionic Morpholino backbone, we show here that Nucleofection® can be used as an
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Evaluation of PARP Inhibitors: Performed on BMG LABTECH’s FLUOstar Omega
BMG LABTECH
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
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Dual Luciferase Reporter (DLR) Assay Certification on the Omega Series of Readers
BMG LABTECH
The Dual-Luciferase® Reporter Assay or DLR™ from Promega is widely used to study gene transcription and regulation. The DLR assay is a two step reaction that uses two luciferase enzymes, from Firefly and Renilla. The Omega series of microplate readers from BMG LABTECH has been granted DLReady™ certification based on the results published in this application note. Two onboard reagent injectors with simultaneous measurement allow for instantaneous detection of both luciferases.
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Investigation of the stereoselectivity of an anti-amino acid antibody utilizing tryptophan fluorescence
BMG LABTECH
The binding sites of proteins such as antibodies are known to often
contain tryptophan (Trp) residues, whose fluorescent properties may be
altered upon ligand binding. Conformational changes within the binding
site or simply the presence of the ligand can result in either fluorescence
quenching or enhancement, which may be utilized to quantitatively
investigate protein-ligand interactions.
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Genomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Tissues through the use of Whole Genome Amplification (WGA)
Sigma Aldrich
Whole Genome Amplification of Formalin-Fixed Paraffin Embedded (FFPE) Tissues
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Analysis of Transferrin using a SMIL coating on deltaDOT’s CE platform
deltaDOT
isoforms including CDT were successfully analysed and resolved using the Peregrine High Performance Capillary Electrophoresis system and deltaDOT’s proprietary LFII™ technology, demonstrating its ability to produce high quality reproducible data in terms of peak mobility (<0.5 % R.S.D.) and peak area (~ 2 % R.S.D.)
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Analysis of Recombinant Human Erythropoietin on deltaDOT’s CE platform
deltaDOT
Recombinant Erythropoietin (EPO) has uses in legitimate treatment of disease such as anaemia, but has become notorious for its use in blood doping in endurance sports. The glycoform fingerprint obtained by this analysis, using the Peregrine High Performance Capillary Electrophoresis system and deltaDOT’s proprietary LFII™ technology, gives high quality reproducible data in terms of peak mobility (< 2 % R.S.D.) and peak area (<2 % R.S.D.). This can be potentially used to identify individual vendo
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Indirect UV Analysis of Amino Acids using by Capillary Zone Electrophoresis with LFII™ Detection
deltaDOT
A mixture of amino acids was successfully analysed using deltaDOT’s Peregrine HPCE at high pH to fully ionize each of the amino acid residues. Indirect UV detection was employed to visualise the 9 amino acids. Excellent resolution has been demonstrated between all 9 amino acid residues, with high reproducibility for migration time (<0.5% R.S.D.) and peak area (<2% R.S.D.). deltaDOT’s core technology of Label Free Intrinsic Imaging, (LFII™) may be applied to indirect UV measurements fo
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A global abstract analysis of selected histone-modifying enzymes using AKS
Active Motif
A global abstract analysis (GAA) was performed on twelve histone-modifying enzymes (ASH1, CARM1, CBP, CKII, EZH2, MSK1, MST1, NSD1, PCAF, RIZ1, SIRT2 and TIP60) to demonstrate how this approach can help uncover biological relationships, scientific information, and overall publication trends from PubMed abstracts and related biological databases. This GAA study showed that published research of histone-modifying enzymes has increased significantly since 2001 and that many reported studies have fo
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Efficient PCR from whole blood using Finnzymes High Performance PCR solution
Labtech International
Finnzymes have developed a range of reagents, consumables and instrumentation to allow fast PCR in inhibiting conditions. Here we describe an application for amplifying exogenous and human genomic DNA from whole blood without the need for DNA purification, and use of special buffers or complicated protocols.
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Fluorescence Polarization Discriminates Green Fluorescent Protein from Interfering Autofluorescence
BMG LABTECH
In recent years, Green fluorescent protein (GFP) has been widely adopted as a versatile marker for reporting on gene expression in a wide range of cells and organisms. However, fluorescence measurements of GFP (unless it is very highly expressed) are invariably contaminated with endogenous autofluorescence from the cells or media. In this application note we introduce a method to discriminate GFP from interfering autofluorescence by using a fluorescence polarization method.
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Invitrogen’s LanthaScreen™ TR-FRET Tyrosine Kinase Assay Using the PHERAstar
Invitrogen Corporation and BMG LABTECH
Several tyrosine kinases were characterized using Invitrogen´s LanthaScreen™ assay. The LanthaScreen™ TR-FRET platform combines terbium as the donor group and fluorescein as the acceptor species.BMG LABTECH’s PHERAstar microplate reader provides the ideal platform to simplify assay development. With its dual wavelength emission detection and five photomultiplier tubes, the PHERAstar provides the speed and sensitivity needed to take full advantage of Invitrogen’s LanthaScreen™ technology.
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Invitrogen’s LanthaScreen™ TR-FRET Protein Kinase C Assay Using the PHERAstar
Invitrogen Corporation and BMG LABTECH
BMG LABTECH’s PHERAstar microplate reader provides the ideal platform to simplify assay development. With its dual wavelength emission detection and five photomultiplier tubes, the PHERAstar provides the speed and sensitivity needed to take full advantage of Invitrogen’s LanthaScreen™ technology.
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Simultaneous Dual Emission Detection for Fast Kinetic BRET Assays
BMG LABTECH
Bioluminescence Resonance Energy Transfer (BRET) is a system for monitoring intermolecular interactions in vivo. The BRET2™ demo kit has been used to prove the feasibility of performing a BRET assay on the POLARstar OPTIMA microplate reader. The POLARstar OPTIMA’s internal reagent injectors for 384-well plate format combined with high-end simultaneous dual emission detection offer a unique advantage for fast kinetic assays where simultaneous emission detection at two wavelengths is required.
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Promega’s Multiplexed Luciferase Reporter and Cell Viability Assays performed on the PHERAstar
BMG LABTECH and Promega Corporation
By multiplexing a reporter assay (EnduRen™, ViviRen™) with a cell viability assay (CellTiter-Glo®), it is possible to determine if reporter response variations are due to changes in cell number and health. In this poster, we demonstrate the combination of several Promega cell-based assays multiplexed in both low-volume 384- and 1536-well plate formats using the BMG LABTECH PHERAstar microplate reader to record luminescence.
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Dynamic Detection
febit biotech gmbh
In contrast to many microarray platforms, the Geniom technology provides the user with a unique feature that allows specific measurements and quality control via real-time monitoring on the chip: the dynamic detection of hybridization and amplification. Presenting this novel measuring principle, Geniom enables advanced applications and new approaches.
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microRNA Profiling
febit biotech gmbh
febit’s Geniom Technology addresses the special requirements for the analysis of miRNA expression, its most significant advantage being the extraordinary adaptability to up-to-date information. Keeping the pace with ever changing updates of the miRNA databases as well and easy to handle adjustments of microarray setups, enable modern analyses of miRNA profiles.
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Genotyping
febit biotech gmbh
The Geniom® technology for mutational and comparative sequence analysis is based on hybridization of sample DNA to specific probes on the custom array. Use genomic sequence information derived from private or public databases, or select fully validated sequences from our pre-selected probe compositions to determine design and setup of your array.
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Validation of TNFα Antibodies with UNIchip® AV - 400
Protagen AG
Protagen AG has developed the UNIchip® family of protein biochips which is extremely useful in providing valuable information for antibody development. The UNIchip® is able to provide the information to identify superior antibody development candidates at an early stage thus saving time and money.
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Analysis of a Complex Sample of Bacterial Whole Cell Lysate from Haemophilus ducreyi using the Finnigan LXQ
Thermo Fisher Scientific
This application determines the feasibility of analyzing a complex mixture, represented by a whole cell lysate of H. ducreyi, on the LXQ linear ion trap using a simple, 1D-SDS PAGE desalting and protein fractionation procedure, and to study the effect of non-MS parameters, such as sample complexity and chromatographic resolution on the number and quality of the spectra generated and the resulting proteins identified, with particular emphasis on membrane and high molecular weight proteins.
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