Navigation Links
A highly sensitive ATP-luminescence assay using VICTOR2 with the dispenser option

INTRODUCTION

In high sensitivity flash type luminescence assays the measurement is done just after dispensing the activating reagent or after a short delay time. Over the past years luminescence substrates have been improved towards providing more stable signal. These so-called glow type substrates allow microplates to be counted without dispensors. However, flash type reagents will always have one advantage, sensitivity. In the glow type the luminescence signal is spread over a wide time scale (for example half life 30 min) and only a portion of it is read (1 - 10 s / well), so a lot of light is not detected. In flash assays most of the light is counted (half-life 5 s).

Adenosine triphosphate (ATP) is the energy reserve essential for viable cells. The concentration of ATP is stable in cells. Therefore the amount of ATP in the well correlates with the number of viable cells. ATP assays can be used for cell proliferation, cytotoxicity and cell adhesion studies. One application is the detection of microbial contamination in milk. In this study ATP samples were assayed with a Boehringer Mannheim flash type kit using VICTOR2 dispenser option. The detection is based on a light emitting (560 nm) reaction of ATP and luciferin in the presence of luciferase enzyme and oxygen.

METHODS

The assay of an ATP dilution series in a plate with a constant luciferase concentration was performed according to the kit instructions (ATP Bioluminescence Assay kit HS II, Boehringer Mannheim, Germany). 50 μL of Luciferase reagent was dispensed into white microplates (Combiplate, Labsystems, Finland) with 25 μL of ATP sample and 25 μL of cell lysis reagent. The wells were measured for 10 seconds after a 1 s delay.

RESULTS

The dilution series proved to be linear over 5 decades with a detection limit of 1.0E-11 M ATP
'"/>

Source:


Page: All 1 2

Related biology technology :

1. Eppendorf Electroporation Application Notes Your participation is highly welcome !!!
2. Eppendorf Electroporation Application Notes Your participation is highly welcome !!!
3. Screening for β2-adrenergic receptor agonists using the pH-sensitive dye,CypHer5, and the IN Cell Analyzer 3000
4. 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
5. The combination of monolithic columns and modern HPLC/MS/MS instrumentation for high chromatographic throughput and sensitive detection of beta-agonists
6. Enzyme Immunoassay for Studying Intracellular Levels of cAMP
7. LiquiChip Kits for bead-based cytokine and kinase assays
8. Quality control of antibodies using the 2100 bioanalyzer and the Protein 200 Plus assay
9. High-content screening of GPCR activation with MetaXpress, AcuityXpress and the Transfluor assay system
10. Bio-Plex Cytokine Immunoassays and ELISA: Comparison of Two Methodologies in Testing Samples From Asthmatic and Healthy Children, Rev A
11. Principles of Curve Fitting for Multiplex Sandwich Immunoassays, Rev B
Post Your Comments:
(Date:1/15/2014)... (PRWEB) January 15, 2014 Freeslate, ... solutions, today announced that Lupin Limited, one of India’s ... CM Protégé PharmD System for high throughput ... India, is focused on a wide range of quality, ...
(Date:1/14/2014)... (PRWEB) January 14, 2014 Histogen, Inc., ... products of cells grown under simulated embryonic conditions, today ... agreement with Suneva Medical, Inc. for physician-dispensed aesthetic products ... , This agreement is an amendment to the ...
(Date:1/14/2014)... BETHESDA, Md. , Jan. 14, 2014  Organovo announced ... from the National Institutes of Health (NIH) to help ... effective treatments to patients on a faster timeline. Organovo ... Sciences (NCATS) and the National Eye Institute (NEI) to ...
(Date:1/14/2014)... OTTAWA, Ontario , Jan. 14, 2014   Kinaxis ®, ... ( SCM ) and sales and operations planning ( S&OP ... the Biomanufacturing Summit , which will be held at ...  At the conference, join Kinaxis customer Elisabeth Kaszas , ...
Breaking Biology Technology:Lupin Selects Freeslate’s CM Protégé PharmD System to Accelerate Polymorph Screening for Drug Development 2Lupin Selects Freeslate’s CM Protégé PharmD System to Accelerate Polymorph Screening for Drug Development 3Histogen and Suneva Medical Expand License for Cell Conditioned Media-based Aesthetic Products Internationally 2Histogen and Suneva Medical Expand License for Cell Conditioned Media-based Aesthetic Products Internationally 3Organovo Announces Collaboration with National Institutes of Health 2Organovo Announces Collaboration with National Institutes of Health 3Event Alert: Kinaxis Customer to Present at the Biomanufacturing Summit "Supply Chain: Improving Network Effectiveness" 2
... Tumors Associated ... with HER2-Positive Breast Cancer, SAN ANTONIO, Texas, Dec. ... a Phase II study on its,first-in-class, oral small molecule HER2 ... the combination of,TYKERB plus capecitabine (Xeloda(R)) showed a reduction in ...
... - Vasogen Inc.,(NASDAQ: VSGN ; TSX:VAS) and ... pharmaceutical and medical devices company, today,announced the ... System,in Germany. Celacade, a first-in-class therapy, is ... targets the destructive chronic,inflammation underlying the development ...
... Mednet, SWOG Will Enhance Quality and Speed Image Transfer at ... Cost While Ensuring 21 CFR Part 11 ... largest,diagnostic imaging network, today announced it has been selected by,Southwest ... trials cooperative groups in the United States, to provide a,secure, ...
Cached Biology Technology:GlaxoSmithKline Announces Data on TYKERB(R) (Lapatinib) Plus Capecitabine for the Treatment of HER2 Positive Breast Cancer Brain Metastases 2GlaxoSmithKline Announces Data on TYKERB(R) (Lapatinib) Plus Capecitabine for the Treatment of HER2 Positive Breast Cancer Brain Metastases 3GlaxoSmithKline Announces Data on TYKERB(R) (Lapatinib) Plus Capecitabine for the Treatment of HER2 Positive Breast Cancer Brain Metastases 4GlaxoSmithKline Announces Data on TYKERB(R) (Lapatinib) Plus Capecitabine for the Treatment of HER2 Positive Breast Cancer Brain Metastases 5GlaxoSmithKline Announces Data on TYKERB(R) (Lapatinib) Plus Capecitabine for the Treatment of HER2 Positive Breast Cancer Brain Metastases 6Vasogen and Grupo Ferrer Announce Initial Roll-out of Celacade - New Therapy for Chronic Heart Failure 2Vasogen and Grupo Ferrer Announce Initial Roll-out of Celacade - New Therapy for Chronic Heart Failure 3Vasogen and Grupo Ferrer Announce Initial Roll-out of Celacade - New Therapy for Chronic Heart Failure 4Vasogen and Grupo Ferrer Announce Initial Roll-out of Celacade - New Therapy for Chronic Heart Failure 5AG Mednet to Provide Image Network for Southwest Oncology Group 2
(Date:4/21/2014)... lower cholesterol can reduce heart attack and stroke risk, ... save the vision of diabetics, scientists say. , In ... of niacin to decrease bad cholesterol levels and raise ... for these patients, said Dr. Pamela Martin, biochemist at ... , Martin is Principal Investigator on a new ...
(Date:4/20/2014)... 20, 2014) A team of researchers from ... College, and Brandeis University has devised a wholly ... involving the so-called retromer protein complex. Retromer plays ... protein (APP) away from a region of the ... toxic byproduct amyloid-beta, which is thought to contribute ...
(Date:4/20/2014)... Although doctors have long known that people with Down ... leukemia (ALL) during childhood, they haven,t been able to ... Institute investigators has uncovered a connection between the two ... the journal Nature Genetics , the researchers track ... abnormality in Down syndrome to the cellular havoc that ...
Breaking Biology News(10 mins):Scientists target receptor to treat diabetic retinopathy 2Scientists target receptor to treat diabetic retinopathy 3'Chaperone' compounds offer new approach to Alzheimer's treatment 2'Chaperone' compounds offer new approach to Alzheimer's treatment 3Dana-Farber researchers uncover link between Down syndrome and leukemia 2Dana-Farber researchers uncover link between Down syndrome and leukemia 3
... La Jolla, CA - Salk Institute professor Terrence J. ... the neural networks revolution in computing in the 1980s, ... of Sciences. The Academy made the announcement today during ... the Academy recognizes distinguished and continuing achievements in original ...
... CARSON CITY, Nev. , April 27 State of ... by participating in The Prevention Plan™, a U.S. Preventive Medicine® program designed to ... before they progress. The program is projected to save Nevada ... , , ...
... , ATLANTA , April 27   Georgia Power received approval April 20 ... more solar options. , , , ... , , ... to include the following: , , , ...
Cached Biology News:Salk scientist Terrence Sejnowski elected to National Academy of Sciences 2U.S. Preventive Medicine Helps Participants of the State of Nevada Public Employees' Benefits Program (PEBP) Become Healthier 2U.S. Preventive Medicine Helps Participants of the State of Nevada Public Employees' Benefits Program (PEBP) Become Healthier 3Georgia Power's Green Energy Program Redesigned to Include More Solar Power 2
... advanced protein cross-linking techniques are defined by ... Shan S. Wong. This valuable hardcover guide ... and current application references. More than 300 ... cross-linking and conjugate preparation from selection ...
... The proteomic service includes protein separation ... using the PDQuest software package from ... mass spectrometry. In order to meet ... preparation, data analysis and format of ...
... provides reagents for immune complex disruption (ICD) ... samples using a combination of low pH ... and transferred to microplate wells which are ... antibody to HIV-1 p24. The immobilized monoclonal ...
... endotoxin-free plasmid DNA preparations are prepared ... quantities from 1mg to multi-gram scales. ... customer needs, and glycerol stocks are ... Transformation, clonal selection (Clonal selection includes ...
Biology Products: