CEP

New Kit Generates Exceptionally Pure Total RNA for RT-PCR

RT-PCR miniprep kit Stratagene Stratagene has determined that standard RNA purification kits are not satisfactory for reverse transcribed PCR (RT-PCR) applications. We...

Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling

Danny Q. Hoang Michael Browning Xinping Yang PeterPingerelli Stratagene Cloning Systems, Inc. Stratagene introduces affinity purified antibodies for NMDA receptorsubunits that are useful for studying expression of the NR2A, NR2B and NR2Csubunit proteins...

Detecting Gastrin-Releasing Peptide Receptor by in situ PCR on Archived Tissue

In situ Adapter for the RoboCycler 40 Temperature Cycler allows rapidamplification of rare mRNA Department of Medicine, University of Illinois at Chicago Figure 1: RoboCycler 40 Temperature Cycler In Situ Adapter Syst...

pFB-ERV: Retroviral Delivery of the Ecdysone Receptor Proteins

Faster, easier screening for difficult transfections when using thecomplete control kit Stratagene We describe the vector pFB-ERV, an MMLV-based replication-defectiveretroviral vector for delivery of the ecdysone receptor proteins RXR and VgEcR.In stable cell lines infected with an estima...

Gene-Specific Primers for NMDA Receptor Subunits

Robert Buchner Peter L. Pingerelli Michael D. Browning Mary Ellen Simcox Stratagene Cloning Systems, Inc. Stratagene is now offering gene-specific primer sets for the rat and humanNMDA receptor. In this article, we describe using these primers in reversetran...

TKB1 Cells Identify Receptor Tyrosine Kinase Interacting Proteins

Institute for Molecular Biology and Biotechnology, McMaster University,Ontario, Canada GST-fusion proteins expressed in TKB1 bacteria are used to affinity purifycellular proteins, which interact with a specific autophosphorylation site onthe Erb...

Quantification of the Purinergic Receptor P2X3 using ICAT and Orthogonal 2D LC-MS/MS with an Ion Trap Mass Spectrometer

David Barnidge , Gargi Choudhary Neuromics, Inc., Minneapolis, MN Thermo Finnigan, San Jose, CA The data presented here was acquired using the Proteome...

Isolation of Low Molecular Weight Digestion Products of the Human Platelet Thromboxane A2 Receptor, Rev A

Joseph W. Turek and Guy C. Le Breton, Department of Pharmacology,University of Illinois at Chicago, College of Medicine, Chicago, IL 60612.Address correspondence to: Joseph W. Turek, 835 South Wolcott, Departmentof Pharmacology M/C 868, University of Illinois at Chicago, Chicago, IL 60612.E-mail: jturek1@icarus.uic.edu Introduction...

Receptor Binding Assays using Fluorescence Polarization Detection: the [FP]2TM Characterized Tracers

. The process is typically highly automated and can provide screening rates over 100,000 samples in a 24 hour period. To achieve this, assays must be simple, p...

Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets.

Recent advances in combinatorial chemistry and genomics programs have led to an unprecedented increase in both the number of compounds and potential drug targets that can be screened. The development of automated liquid h...

Ca++-Induced G Protein Coupled Receptor (GPCR) Activation Monitored via Aequorin Luminescence in the LMax Microplate Luminometer (MaxLine Application Note #42)

Jinfang Liao, M.D., Ph.D. and Evelyn McGown, Ph.D. Molecular Devices Corporation, 8/00 The photoprotein aequorin, a bioluminescent protein from the coelentratejellyfish Aequorea victorea, has proven to be highly sensitive to the trace amount of Ca in cells. The aequorin complex which contains the...

Gq-coupled Receptor Assays Using FLEXstation

Departments of...

Gq-coupled receptor assays using FLEXstation: M1 muscarinic receptor

SIMPLE, RAPID EXPERIMENTAL SET-UP AND DATA ANALYSIS DETERMINE EC EXCELLENT FOR ASSAY DEVELOP-MENT AS DEMONSTRATED BY Z FACTOR INTROD...

Monitoring Receptor-Mediated Changes in [Ca2+]i Using Single-Wavelength and Dual-Wavelength Indicators on the FlexStation Workstation

Simon Lydford & Petra Averbeck, Molecular Devices Limited, U.K. The FlexStation scanning fluorometer integrates sensitive optics, fluid transfer and temperature control, making it ideal for kinetic, cell-based fluorometric assays, such as the measurement of intracellular calcium, membrane potential and intracellular pH. <...

Gq-coupled receptor assays: a comparison of FLEXstation and FLIPR

SIMPLE, FAST EXPERIMENTAL SET-UP Activation of G -coupled...

State's investment in early-stage businesses is changing perceptions

Attorney Joe Hildebrandt has been a part of many deals connecting promising young Wisconsin companies with investors, some close at home and others in the financial centers of the nations East and West coasts. Over the years, Hildebrandt told a legislative committee last week, he's battled the notion that Wisconsin is a "flyover state" that nei...

The development of a CXCR2 chemokine receptorbinding assay using the LEADseeker Multimodality Imaging System

CXCR2 • GPCR • receptor binding assay • LEADseeker • SPA Imaging Beads Chemo-attractant cytokines (chemokines) are low molecular weight, pro-inflammatory cytokines with potent leukocyte activation and/or chemotactic activity (1). Chemokines are classified into four families (CXC, CC, C, and CX...

Performance of Ad-A-Gene EGFP-Glucocorticoid Receptor (EGFP-GCCR): an adenoviral vector gene delivery system

adenoviral vector • viral transduction • EGFP • IN Cell Analyzer • gene delivery There is a requirement in the drug discovery process to investigate lead candidate drugs arising from primary screens and to validate those compounds in suitable biological assays. There is also a need in early-stage drug discov...

The development of a melanocortin MC4 receptor binding assay using the LEADseeker Multimodality Imaging System

Melanocortin • receptor binding assay • LEADseeker • SPA Imaging Beads Pro-melanocortins are present in high levels in the pituitary and are processed into three major peptide hormone fragments: adrenocorticotrophin (ACTH), the melanocyte stimulating hormones, and β-endorphin. The various melanocyte stimulat...

The development of a melanocortin MC5 receptor binding assay using the LEADseeker Multimodality Imaging System

Melanocortin • receptor binding assay • LEADseeker • SPA Imaging Beads Pro-melanocortins are present in high levels in the pituitary and are processed into three major peptide hormone fragments: adrenocorticotrophin (ACTH), the melanocyte stimulating hormones, and β-endorphin. The various melanocyte stimulat...

The development of a calcitonin-gene related peptide (CGRP) receptor-binding assay using the LEADseeker Multimodality Imaging System

CGRP • receptor binding assay •LEADseeker • SPA Imaging Beads Calcitonin-gene related peptide (CGRP) is believed to have an important physiological role in the central nervous and cardiovascular systems. Indeed, using CGRP receptors isolated from coronary and basilar arteries, studies have i...

Screening for potential beta 2-adrenergic receptor antagonists using CypHer5E and IN Cell Analyzer 1000

5, a pH sensitive dye, has shown utility in β2-adrenergic receptor agonist screening (1). CypHer5 has been used to obtain dose-response and rank-order potency data for both agonist and antagonist treatment of β2-adrenergic receptor expressing cells (2). For live-cell receptor internalization studies,...

The use of CypHer5 for receptor internalization studies in both a range of GPCRs and a non-GPCR

CypHer5 is a red-excited, pH-sensitive cyanine dye that is non-fluorescent at pH 7.4 and maximally fluorescent at pH 5.5. It is therefore ideally suited for monitoring internalization of cell surface receptors via acidic endosomal vesicles upon agonist stimulation (1, 2). <P >In the assay, the receptor can be epitop...

The use of CypHer5E and the IN Cell Analyzer 1000 for live-cell receptor internalization studies

Measurement of receptor internalization using the pH-sensitive cyanine dye, CypHer™5, has previously been reported (1,2,3). An enhanced version of this dye that shows greater relative fluorescence intensity when maximally fluorescent, greater solubility, and stability has now been developed. The enhanced dye, CypHer5E, is a red-...

Screening for β2-adrenergic receptor agonists using the pH-sensitive dye,CypHer5, and the IN Cell Analyzer 3000

High-throughput screening for potential targets is a major requirement in drug development and a key application is in the area of receptor activation, particularly G-protein coupled receptors (GPCRs) (1). <P >It is estimated that there are still 100–200 orphan GPCRs of potential therapeutic value requiring identification of...

Development of a histamine H1 receptor binding assay using the LEADseeker Multimodality Imaging System

receptor • GPCR • receptor binding assay • LEADseeker • SPA Imaging Beads Histamine is a biogenic amine that plays important pathophysiological roles in central and peripheral tissues. Generally released from stimulated mast cells, it contributes to several biological processes that characteri...

Development of a sensitive enzyme fragment complementation assay for cyclic adenosine 3', 5' monophosphate and its validation for pharmacological screening at G protein-coupled receptors

G protein-coupled receptors (GPCRs) are the largest family of transmembrane receptors and are responsible for the transduction of a diverse range of extracellular signals. Up to 40% of clinically ma...

Development of an NK2 neuropeptide receptor binding assay using LEADseeker Multimodality Imaging System

Neurokinins, also known as tackykinins, are a family of small neuropeptides. Members of this family include peptides, substance P (SP), neurokinin A (NKA), neurokinin B (NKB), and the recently discovered hemokini...

The development of a somatostatin sst4 receptor-binding assay using the LEADseeker Multimodality Imaging System

Somatostatin (SS) is a neurotransmitter/hormone with a wide range of biological actions (1–3) and a reduction in cortical SS levels has been reported in Alzheimer’s and Parkinson’s diseases. The SS analog...

The development of a serotonin 5-HT2C receptor-binding assay using the LEADseeker Multimodality Imaging System

In mammals, serotonin (5-hydroxytryptamine, 5-HT) is a regulator of neural transmission which induces its biological effect by binding to serotonin receptors. <P >The serotonin 5-HT rece...

Development of an adrenergic α2a receptor binding assay using the LEADseeker Multimodality Imaging System

The α +...

Development of a CRF1 neuropeptide receptor binding assay using LEADseeker Multimodality Imaging System

In the mammalian brain, the corticotrophin-releasing factor (CRF) system comprises four CRF-related peptides (CRF and urocortin 1, 2, and 3) and two G protein-coupled CRF receptor subtypes (CRF...

Cell lines expressing recombinant aequorin and a G-protein coupled receptor for functional screening

Many G-protein coupled receptors (GPCRs) trigger, upon binding of an agonist, a transient increase in intracellular calcium concentration. This variation acts as an internal secondary messenger and is an important modulator of many physiological mechanisms (reviewed by Rink (1990), Tsunoda (1993) and by Santella & Carafoli (1997)). Measure...

Use of FlashPlate Technology for In Vitro Measurement of [35S]-GTPγS Binding in CHO Cells Expressing the Human 5-HT1B Receptor

SmithKline Beecham Pharmaceuticals, UK FlashPlate is a 96-well white polystyrene microplate with scintillant-coated wells designed for highvolume, in-plate radiobinding assays. This study investigates the use of FlashPlate as a platform for the measurement of G-protein act...

Use of Novel FlashPlate Technology to Measure cAMP Accumulation in Chinese Hamster Ovary Cells Expressing Human -2 Adrenoreceptors

SmithKline Beecham Pharmaceuticals, UK Due to recent technologies in the pharmaceutical industry that have resulted in an increase in the production of potential drug candidates, it has become vital that screening assays are high throughput. FlashPlate technology from PerkinElmer Life Sciences has pro...

Use of FlashPlate Technology for In Vitro Measurement of 125I-Labeled TGF-1 Binding on Chimeric Extracellular Domain of Type II Transforming Growth Factor Receptor

Dpartement dImmunologie, Laboratoires Fournier S.A., France TGF- type I and type II receptors form ligand dependent heteromeric signaling complexes, in which transforming growth factor- receptor type II (TRII) tends to act as the primary receptor....

Determination of Acepromazine and its Major Metabolite in Equine Serum by LC-MS/MS using the Finnigan LCQ Deca XP Plus Ion Trap Mass Spectrometer

Wayne Skinner and Scott Stanley, University of California, Davis <p...

Biologist who cloned Dolly the sheep believes therapeutic cloning will gain acceptance

Wilmut, a biologist with the <a href="http://www.re...

Delivering value instead of advertising - what a concept!

We find that families do not want to have their...

Misconceptions dominate the immigration debate

A recent arti...