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

Costar™ solid white 384-well NBS™ coated microplate (Corning)

Protease-free BSA (Fluka)

EDTA-free protease inhibitor cocktail tablets (Roche)

Buffer: 25 mM Hepes pH 7.4
5 mM MgCl2
1 mM CaCl2
0.2% BSA (w/v) plus protease inhibitor cocktail

GraphPad Prism™ software v4.0 (GraphPad Software)


Protocol
Human recombinant CRF1 receptor membrane preparation was used in conjunction with [125I]Tyr0-CRF ligand and WGA PS SPA Imaging Beads. Non-specific binding (NSB) was determined in the presence of 200 nM urocortin. The standard assay format was as follows:

1) Reagents were added in the following order: buffer, unlabeled ligand (NSB wells) labeled ligand, membrane, and bead. Total assay volume was 50 µl.

2) Wells contained 10 µl of 1.5 nM [125I]Tyr0-CRF (final concentration 0.3 nM) unless otherwise stated. Wells also contained 0.3 µg of membrane and 30 µg of beads, which were added in 10 µl volumes.

3) NSB wells contained 10 µl of 1000 nM urocortin (final concentration 200 nM) in addition to the above.

4) Plates were sealed and incubated in darkness for 18 h at room temperature (20–25 °C).

5) Following incubation, plates were imaged on LEADseeker Multimodality Imaging System for 5 min using quasi-coincident averaging and 3 ×3 binning.


Results
Kd determination
Saturation binding was performed with dilutions of [125I]Tyr0-CRF to give a range of concentrations from 0.06 to 4.96 nM in the wells. Figure 1 shows the saturation curve, which was fitted using non-linear regression with the data analysis s
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. The let-7 microRNA Family Regulates RAS: Implications for Development and Oncogenesis
2. Rapid Method Development With the BioLogic DuoFlow Chromatography System for the Purification of His-Tagged Proteins
3. A Multiple Mutation Model System as a Test Development and Training Tool for Denaturing Gradient Gel Electrophoresis
4. Development of a Multiplex Bead-Based Assay for Antibody Screening of a Nonhuman Primate Colony on the Bio-Plex System
5. Development of a histamine H1 receptor binding assay using the LEADseeker Multimodality Imaging System
6. 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
7. Development of an NK2 neuropeptide receptor binding assay using LEADseeker Multimodality Imaging System
8. Development of an adrenergic α2a receptor binding assay using the LEADseeker Multimodality Imaging System
9. Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets.
10. The development of a CXCR2 chemokine receptorbinding assay using the LEADseeker Multimodality Imaging System
11. The development of a melanocortin MC4 receptor binding assay using the LEADseeker Multimodality Imaging System
Post Your Comments:
(Date:6/2/2015)... , June 2, 2015   Hospira, Inc. ... today announced that its partner, Celltrion – a global biopharmaceutical ... National Health Surveillance Agency in Brazil ... antibody approved for use in Brazil ... also markets and sells biosimilar infliximab in 26 European ...
(Date:6/2/2015)... , June 2, 2015   Sigma-Aldrich Corporation ... and Technology company, announced today it has entered ... to launch genetically engineered human neural differentiated cells ... research of neurological diseases. These new products, in ... and versatile tools for the study of central ...
(Date:6/2/2015)... 2015 Through rare discovery of new ... Corporation is enabling World Scientists to actually see, ... sample, displaying the full nature (position, shape, size, number) ... Jeff Porter, Co-Founder of the Yancy Corporation, says, "Our ... rare. They come once in a lifetime, if at ...
(Date:6/1/2015)... , June 1, 2015  Turing Pharmaceuticals ... Eliseo Oreste Salinas , MD, MSc,  has joined ... Dr. Salinas, who holds a medical ... brings a record of significant accomplishments in the ... Dr. Salinas has played a leadership ...
Breaking Biology Technology:ANVISA Approves First Infliximab Biosimilar in Brazil 2ANVISA Approves First Infliximab Biosimilar in Brazil 3ANVISA Approves First Infliximab Biosimilar in Brazil 4ANVISA Approves First Infliximab Biosimilar in Brazil 5Sigma-Aldrich to Launch XCell Science Genetically Engineered iPSC-Derived Neural Disease Cells 2Sigma-Aldrich to Launch XCell Science Genetically Engineered iPSC-Derived Neural Disease Cells 3Scientists Eying Rare Discovery Of New Physics & Light Microscope Technology 2Turing Pharmaceuticals Names Eliseo Salinas, MD and MSc, President of Research and Development 2Turing Pharmaceuticals Names Eliseo Salinas, MD and MSc, President of Research and Development 3
... Technology Network is committed to helping improve patient safety ... We salute the administration and the efforts of Secretary ... health technology. If you are interested in learning ... event. , ,The University of Wisconsin and ...
... stories critical of the ethics of contemporary business executives ... the academic literature. A noteworthy exception is "Beyond Selfishness ... the Fall 2002 issue of the Sloan Management Review ... an elegantly argued essay that should be required reading ...
... its MOVEit family of secure file-transfer software products now ... server file delivery. The new features include transfer persistence, ... and MOVEit clients now provide transfer persistence by retrying ... Files that fail at the beginning or end of ...
Cached Biology Technology:The doctor is in: Thompson prescribes better health information technology 2The doctor is in: Thompson prescribes better health information technology 3The doctor is in: Thompson prescribes better health information technology 4Is your competitive edge really a wedge? 2Is your competitive edge really a wedge? 3
(Date:4/27/2015)... more than four decades, AUVSI,s Unmanned Systems ... event for the unmanned land, sea, and air industries. ... local, national, or trade news organization interested in gathering ... applications of unmanned technologies. The conference will explore the ... and how it will soon impact the lives of ...
(Date:4/20/2015)... 20, 2015 Huntington Memorial Hospital is the ... a new miniaturized, wireless monitoring sensor to manage heart ... and only FDA-approved heart failure monitoring device that has ... by physicians to manage heart failure. The ... in the pulmonary artery (PA) during a non-surgical procedure ...
(Date:4/14/2015)... 2015 NXT-ID, Inc. (NASDAQ: NXTD ) ... on the growing mobile commerce market, announces that its ... New York Design Awards under the category  ,Product Design ... Design Awards are part of a global multi-disciplinary awards ... from the marketplace, industry and judging panel. Winners will ...
Breaking Biology News(10 mins):UNMANNED SYSTEMS 2015: Largest International Conference and Trade Show for the Drone and Unmanned Technology Industry 2UNMANNED SYSTEMS 2015: Largest International Conference and Trade Show for the Drone and Unmanned Technology Industry 3UNMANNED SYSTEMS 2015: Largest International Conference and Trade Show for the Drone and Unmanned Technology Industry 4UNMANNED SYSTEMS 2015: Largest International Conference and Trade Show for the Drone and Unmanned Technology Industry 5Huntington Memorial Hospital Becomes First in the San Gabriel Valley to Offer Patients a New Heart Failure Monitoring Solution 2Huntington Memorial Hospital Becomes First in the San Gabriel Valley to Offer Patients a New Heart Failure Monitoring Solution 3NXT-ID's Wocket Smart Wallet Nominated for 2015 New York Design Awards in 'Product Design- Technology' 2NXT-ID's Wocket Smart Wallet Nominated for 2015 New York Design Awards in 'Product Design- Technology' 3
... the brain than previously thought, a new study shows. ... first detected by part of the brain called primary visual ... head where visual information first enters the cortex of the ... in animals but has not been tested on human beings ...
... (September 5, 2007) King Abdullah University of Science ... Cairo (AUC) signed a Memorandum of Understanding on August ... development that will positively impact the Kingdom of Saudi ... president of KAUST, and Dr. David D. Arnold, president of ...
... of Engineering may have discovered the path toward ... bacteria that have developed resistance to common antibiotics. ... of events occurring in bacteria that opens to ... Currently, three classes of bactericidal antibiotics are used ...
Cached Biology News:Color contrast is 'seen' by the brain early doors 2KAUST and American University in Cairo to collaborate on research and academic development 2Boston University biomedical engineers find chink in bacteria's armor 2Boston University biomedical engineers find chink in bacteria's armor 3
... xi cell, 20 cm, is used for vertical ... in agarose gels. The cell is supplied without ... cooling core with gaskets, lower buffer chamber, lid ... sandwich clamps, upper buffer dam, casting stand with ...
... is tailored to highly enrich ... of non-human primate (Rhesus monkey) ... the StemSep compatible 0.5 Tesla) ... CD4+ T cell content of ...
... Adept CE 4100 high pressure dual piston pump ... detector for added flexibility. The system ... interface unit, CE 4901., The CE 4901 PowerStream ... used with Windows 98 and 2000, offers the ...
Anti 4-hydroxy-2-hexenal (4-HHE) Monoclonal Antibody (HNEJ-2)...
Biology Products: