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)

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.

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


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:10/8/2015)... -- The ALS Association, in partnership with Prize4Life, is pleased ... communication technology solutions for people living with ALS. ... lateral sclerosis) is a progressive neurodegenerative disease that affects ... cord. Eventually, people with ALS lose the ability to ... total paralysis and death within two to five years ...
(Date:10/8/2015)... ... October 08, 2015 , ... ProMIS Neurosciences, Inc. (TSX: ... develop and commercialize intellectual property rights belonging to The University of British Columbia ... affords exclusive worldwide rights to intellectual property based on use of the ProMIS™ ...
(Date:10/7/2015)... Worth, Texas (PRWEB) , ... October 07, 2015 ... ... Cell Institute ( ) will present a public educational seminar on the ... on Saturday, October 10th from 1:00 pm – 3:00 pm at the Hilton ...
(Date:10/7/2015)... Oct. 7, 2015   VWR (NASDAQ: ... laboratory products, services and solutions, announced today that ... specialty solvent company that performs high volume purification ... enhance VWR,s growing laboratory and production chemical manufacturing ... manufacturing customers. Manuel Brocke-Benz , President ...
Breaking Biology Technology:
... November 13 Axelar AB today announced that,promising ... concerning their,lead anticancer product AXL1717, an insulin-like growth ... at the AACR-NCI-EORTC,International Conference on Molecular Targets and ... being conducted in Sweden,on advanced-stage cancer patients with ...
... , SOUTH SAN FRANCSICO, Calif., Nov. 12 ... encompassing a completely novel mechanism for generating massive ... sequences. The technology platform is referred to ... issued in both the United States (#7,585,957) and ...
... 12 Neogen Corporation (Nasdaq: NEOG ) has been selected ... Friday, Nov. 13, to celebrate Neogen,s 20th anniversary of being listed ... on behalf of the countless many who have made Neogen,s success ... has come a long way in the 20 years since our ...
Cached Biology Technology:
(Date:9/24/2015)... , September 24, 2015 ... september 2015 Kerv ( ... finanstjänster, lanserar idag världens första kontaktlösa betalningsring ... in 77 000 GBP för massproduktion via ... ) , Kerv-bärare kan ...
(Date:9/10/2015)... 10, 2015 Report Details ... Selling Opportunities and Revenue Prospects to Help You ... biologics, especially new drug classes? Get the latest ... with exclusive market data and industry knowledge, benefitting ... results, opportunities and sales predictions. Visiongain,s new ...
(Date:9/10/2015)... ATLANTA , Sept. 10, 2015 ... Cleveland Clinic Wellness to create an interactive, image-based ... self-service health and wellness kiosk.  The unique assessment ... Age"— a number that suggests an individual,s biological ... biometric values as measured by the kiosk. ...
Breaking Biology News(10 mins):
... too much water, including sports drinks, when exercising, ... in prolonged types of endurance exercise at risk ... who study disorders of water metabolism. Such exercise ... This serious condition, known as exercise-induced hyponatremia (EAH), ...
... A new, fast and inexpensive way to mutate genes and ... of Utah geneticists. They tested the method by mutating a ... and limbs in the mutant mouse on the left, compared ... The new method promises to speed efforts to learn more ...
... nanoparticle made in a laboratory at the University of ... device for treating glaucoma, an eye disease that can ... “The nanoparticle can safely get past the blood-brain barrier ... said Sudipta Seal, an engineering professor with appointments in ...
Cached Biology News:
Recalibration of HL-2000-CAL products...
Mature rat whole blood (Sprague-Dawley, averaging 7-8 weeks of age, either sex) is collected at the time of slaughter and processed within 45 minutes. It is then filtered through gauze, pooled in an ...
Several Pichia pastoris strains are available to allow you to optimize expression and recovery of your protein of interest. Table 1 provides information to help you choose the appropriate strain....
Biology Products: