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

medium with 10% fetal bovine serum, 1% Glutamine Pen-Strep and 400 g/mL G418.

2. The day before the experiment, cells in growth medium as above were seeded in 96-well microplates at a density of 60,000 cells/well and incubated overnight in a 37C, 5% CO2 incubator.

3. Cells were loaded with coelenterazine by aspirating the growth medium from the wells and replacing it with 100 L coelenterazine loading solution/well. They were then incubated at 37C with 5% CO2 for an additional 2 hours.


ASSAY PROCEDURE
1. A dilution series of UTP was prepared in H&H with 1% BSA concentrations ranging from 0.2 M to 200 M, with the diluent (0 M UTP) as the blank. All were placed in 15-mL conical tubes.

2. The plate containing the coelenterazine-loaded cells (100 uL/well) was placed into the drawer of the Lmax.

3. The SOFTmax PRO for Lmax was set up to inject 100 L of UTP reagent through the M injector and to begin reading immediately and take 20 consecutive 1-second reads on selected replicate wells.

4. Starting with the blank (0 M UTP in diluent) the M injection system was sequentially filled with increasing concentrations of UTP solutions by pumping 1.5 mL through it and then the selected replicate target wells for each concentration were read. The UTP-containing tubes were placed in a small rack on top of the Lmax so that the injector inlet tubing could easily be moved from one tube to the next higher concentration after each set of measurements was complete.


REACTION PROFILES AND DOSE/RESPONSE RESULTS
The reaction profiles of coelenterazine(h)-loaded cells to 0.1 - 3 uM UTP (final concentrations) are shown in Figure 2. When CHO cells expressing apo-aequorin protein (CHOMQ6 cells) were incubated with 2.5 uM coelenterazine(h), then exposed to UTP (a potent purinergic agonist), they responded
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. A New C-Terminal GST Vector for Protein Production in S. pombe
2. New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
3. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
4. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
5. New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
6. Optimized Imaging of Protein Gels Stained with Coomassie Brilliant Blue Dye
7. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
8. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
9. Yeast Protein Production System Features High Yields and One-Step Purification
10. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
11. Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence

Post Your Comments:
(Date:11/5/2009)..., November 5, 2009 at 4:30 p.m. Eastern Time ,,...stCall/ - OncoGenex Pharmaceuticals, Inc. ("OncoGe...rted unaudited financial results for the third qua...reviewed the Company,s highlights for the third qu...sults reflect the operations of OncoGenex Technolo...
(Date:11/5/2009)...2009 Senomyx, Inc. (Nasd...y taste receptor-based technologies to discover no... ingredient supply industries, today provided a co...the third quarter ended September 30, 2009. Reven...9, compared to $4.0 million for the third quarter ...
(Date:11/5/2009)...urgery and vision correction experts listed in the...volved at the recently concluded American Academy ...Francisco, California. Dr. Stephen Brint, MD, Dr. ...MD, Dr. Peter Rapoza, MD, Dr. Michael Raizman, MD,...Dr. Edward Manche, MD, and Dr. Parag Majmudar, MD,...
(Date:11/4/2009)...Newswire-FirstCall/ -- Star Scientific, Inc . (...lans to introduce the CigRx(TM) nutraceutical prod...ceuticals, for worldwide marketing and sales in pa...VTIV). inVentiv Health offers a complete range of...e product lifecycle, in a range of healthcare cate...
Breaking Biology Technology:OncoGenex Reports Third Quarter 2009 Financial Results 2OncoGenex Reports Third Quarter 2009 Financial Results 3OncoGenex Reports Third Quarter 2009 Financial Results 4OncoGenex Reports Third Quarter 2009 Financial Results 5OncoGenex Reports Third Quarter 2009 Financial Results 6OncoGenex Reports Third Quarter 2009 Financial Results 7OncoGenex Reports Third Quarter 2009 Financial Results 8SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 2SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 3SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 4SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 5SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 6SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 7SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 8SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 9SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 10SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 11SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 12SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 13SENOMYX ANNOUNCES CORPORATE UPDATE AND THIRD QUARTER 2009 FINANCIAL RESULTS 14Leading Vision Correction Experts in Trusted LASIK Surgeons Directory Educate Fellow Doctors at AAO Annual Meeting 2Leading Vision Correction Experts in Trusted LASIK Surgeons Directory Educate Fellow Doctors at AAO Annual Meeting 3Leading Vision Correction Experts in Trusted LASIK Surgeons Directory Educate Fellow Doctors at AAO Annual Meeting 4Leading Vision Correction Experts in Trusted LASIK Surgeons Directory Educate Fellow Doctors at AAO Annual Meeting 5Leading Vision Correction Experts in Trusted LASIK Surgeons Directory Educate Fellow Doctors at AAO Annual Meeting 6Leading Vision Correction Experts in Trusted LASIK Surgeons Directory Educate Fellow Doctors at AAO Annual Meeting 7Leading Vision Correction Experts in Trusted LASIK Surgeons Directory Educate Fellow Doctors at AAO Annual Meeting 8Leading Vision Correction Experts in Trusted LASIK Surgeons Directory Educate Fellow Doctors at AAO Annual Meeting 9Star Scientific Plans Worldwide Marketing and Sales of CigRx(TM) Nutraceutical in Partnership with inVentiv Health 2Star Scientific Plans Worldwide Marketing and Sales of CigRx(TM) Nutraceutical in Partnership with inVentiv Health 3Star Scientific Plans Worldwide Marketing and Sales of CigRx(TM) Nutraceutical in Partnership with inVentiv Health 4
...ober 6, 2008 at 3:00 p.m. EDT, BROOMFIELD, Colo.,... company developing genetically-targeted therapies...es, today announced that,Richard Brewer, the compa.... Michael Bristow, M.D., ARCA,s chairman and chief...Third Annual JMP Securities Healthcare Focus,Confe...
...erapy in U.S. to be Indicated ...ell Therapeutics, Inc. (CTI),(Nasdaq and MTA: CTIC...ntal,Biologics License Application (sBLA) with the...f Zevalin(R) ([90Y]-ibritumomab tiuxetan) as,conso...iously untreated,patients with follicular non-Hodg...
...No. 40/2008, - To OMX Nordic Exchange Copenhagen ...t of license to Roche Diagnostics for,use of the U...oprietary,LNA(TM) detection technology., Roche Di...alTime ready,qPCR assays with the Universal ProbeL...M) detection technology., "This extension of our ...
Other Biology Technology:ARCA biopharma to Present at JMP Securities Healthcare Focus Conference 2ARCA biopharma to Present at JMP Securities Healthcare Focus Conference 3ARCA biopharma to Present at JMP Securities Healthcare Focus Conference 4ARCA biopharma to Present at JMP Securities Healthcare Focus Conference 5Cell Therapeutics Submits Supplemental Biologics License Application for Zevalin Use in First-Line Consolidation Therapy 2Cell Therapeutics Submits Supplemental Biologics License Application for Zevalin Use in First-Line Consolidation Therapy 3Cell Therapeutics Submits Supplemental Biologics License Application for Zevalin Use in First-Line Consolidation Therapy 4Cell Therapeutics Submits Supplemental Biologics License Application for Zevalin Use in First-Line Consolidation Therapy 5Exiqon A/S Today Announces a Grant of License to Roche Diagnostics 2Exiqon A/S Today Announces a Grant of License to Roche Diagnostics 3
(Date:11/5/2009)...of Biology Teachers (NABT) will recognize Leonard ...Naugatuck Valley Community College (NVCC) in Water...on Award during the NABT annual professional devel...n Denver, Colorado. , The Evolution Education Aw...ological Sciences,(AIBS) and Biological Sciences ...
(Date:11/5/2009)... been awarded a training grant in the amount of $2...its Integrative Graduation Education and Research ...grant to be awarded to Kent State. The grant, whic...stment Act of 2009, runs through 2014. , The gra...ent aquatic resource sensing (EARS). The purpose o...
(Date:11/4/2009)...tural science) , United States Using Less Wate...ay than it did 35 years ago, despite a 30 percent ...tly attributable to alternative cooling methods at...s. According to the latest USGS water use report, ... goes to cooling thermoelectric power plants. Irri...
Breaking Biology News(10 mins):Biologists, educators recognize excellence in evolution education 2Kent State receives $2.7 million NSF training grant for environmental aquatic resource sensing 2USGS science picks 2USGS science picks 3USGS science picks 4USGS science picks 5USGS science picks 6USGS science picks 7BioNeutrals Ygiene 28TM 29 Hospital Grade Antimicrobial Proven To Be Completely Inert to Steel In Independent Corrosion Testing 13384 1BioNeutrals Ygiene 28TM 29 Hospital Grade Antimicrobial Proven To Be Completely Inert to Steel In Independent Corrosion Testing 13384 2BioNeutrals Ygiene 28TM 29 Hospital Grade Antimicrobial Proven To Be Completely Inert to Steel In Independent Corrosion Testing 13384 3Radius Investigational Bone Anabolic Agent BA058 Increased Bone Mineral Density 28BMD 29 at Key Fracture Sites in Phase 2 Clinical Trial in Postmen 4829 1Radius Investigational Bone Anabolic Agent BA058 Increased Bone Mineral Density 28BMD 29 at Key Fracture Sites in Phase 2 Clinical Trial in Postmen 4829 2Radius Investigational Bone Anabolic Agent BA058 Increased Bone Mineral Density 28BMD 29 at Key Fracture Sites in Phase 2 Clinical Trial in Postmen 4829 3Radius Investigational Bone Anabolic Agent BA058 Increased Bone Mineral Density 28BMD 29 at Key Fracture Sites in Phase 2 Clinical Trial in Postmen 4829 4While Some New Agents Will Achieve Strong Sales the Migraine Drug Market Will Decline 241 3 Billion by 2018 as Generic Erosion Takes Toll on Top Sel 53712 1While Some New Agents Will Achieve Strong Sales the Migraine Drug Market Will Decline 241 3 Billion by 2018 as Generic Erosion Takes Toll on Top Sel 53712 2While Some New Agents Will Achieve Strong Sales the Migraine Drug Market Will Decline 241 3 Billion by 2018 as Generic Erosion Takes Toll on Top Sel 53712 3
...pert at Johns Hopkins who has spent nearly three d...nfection known as Rocky Mountain spotted fever war...rial disease in Arizona is a growing and dangerous...infectious organisms beyond traditional state boun... England Journal of Medicine, the Hopkins patholog...
...h Institute have found that three bacterial specie... cancer--a discovery the researchers hope will lea...disease. , The findings also open the question of...l role. , The current discovery, reported in the J...that elevated levels of three particular bacteria ...
...nt a great advance for human health. It is based o...NOXIDE system, used in refrigeration towers of fac... be the focus for the proliferation of Legionnaire... team of INASMET-Tecnalia experts designed a refri...olated laboratory (P2 level). A wide range of tria...
Other Biology News:Infectious disease expert warns of spread of Rocky Mountain spotted fever 2Infectious disease expert warns of spread of Rocky Mountain spotted fever 3Forsyth scientists find three bacteria associated with oral cancer 2Forsyth scientists find three bacteria associated with oral cancer 3
Anti-Cytomegalovirus (CMV) Immediate Early Antigen Monoclonal Antibody, Unconjugated, Clone 3G9.2 from CHEMICON
Goat Anti-DRAK 2 Polyclonal Antibody, Unconjugated from Novus Biologicals
Rabbit Anti-Human Secretory Carrier Membrane Protein 2 (SCAMP2) Polyclonal Antibody, Unconjugated from Lifespan Biosciences
... proprietary high-performance desalting resin, exc...g and protein-recovery characteristics compared to...aining as low as 25 g/ml of protein can be process...5% retention of salts and other small molecules ( ...
Biology Products: