Navigation Links
Calcium Flux Assays using the LabChip 3000

d control concentration is measured (Step 6). Assay stability can then be assessed by examining the stability and robustness of the response over time (Step 7).

If the assay is deemed stable, then validation will continue with optimization of throughput and cell usage and confirmation of assay stability over multiple runs (Steps 8 and 9). If the assay is not stable, then optimization of buffer conditions and cell growth conditions will be required prior to validation.

III. Assay Development Steps

1. Choose Cell Line, Cell Growth and Cell Buffer Conditions
A microfluidic assay using the CHO-m1 cell line was developed. Cells were grown in Ham's F12 medium adjusted to contain 1.5 g/L sodium bicarbonate with 100 U/mL penicillin, 0.1 mg/mL streptomycin, 2 mM L-glutamine, 0.1 mg/mL Geneticin G418, 1 mM sodium pyruvate, 10 mM HEPES and 10% Fetal Bovine Serum.

Cells were maintained at 37 oC and 5% CO2 and subcultured in monolayers in T75 flasks using standard cell culture methodologies. Cells were split at 80-90% confluency. The day before each assay cells were seeded at 3 x 106 cells into T75 flasks and incubated overnight at 37 oC and 5% CO2.

2. Label, Suspend and Wash Cells
Cells are labeled at room temperature with two calcium sensitive dyes , Fluo4 and Fura Red, while attached to the plate to minimize cell stress due to multiple pelleting and suspension steps. The minimum concentration of dye that yields bright fluorescently-labeled cells should be used. Typically, 6 x 106 cells are loaded with 6 mL of physiological buffer containing 1 μM Fluo-4 and 1 μM Fura Red. In addition, anion transport inhibitors such as probenecid should be used for cell lines such as CHO-m1 that rapidly export Ca2+ indicators. After loading of dye, the cells are suspended
'"/>

Source:


Page: All 1 2 3 4 5 6 7

Related biology technology :

1. Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal Fibroblasts
2. Cis-Reporting System Monitors Intracellular Calcium Mobilization
3. Cis-Reporting System Monitors Intracellular Calcium Mobilization
4. Calcium Plus Assay Kit, Superior Detection for Specialized Targets
5. FLIPR Calcium Assay Kit Datasheet
6. FLIPR Calcium 3 Assay Kit Datasheet
7. FLIPR Calcium 3 Assay Kit - An Optimal No-Wash Fluorescence Calcium Indicator Reagent for Non-Optimal Calcium Responses
8. HEK293 Calcium Flux Assay
9. Fluorescence-Based Single-Tube Assays to Rapidly Detect Human Gene Mutations
10. Matched siRNAs and Assays: Ambion + Applied Biosystems = RNAi Success
11. Using Validated siRNAs in Functional Genomic Assays
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Calcium Flux Assays using the LabChip

(Date:12/19/2014)... BioPlus Specialty Pharmacy (BioPlus), one of the nation’s ... Pharm.D. to the newly created position of Vice President ... Dr. Maroulis will continue to manage the entire specialty ... pharmacies as the company expands. Dr. Maroulis has been ... has served in many different capacities in the pharmacy, ...
(Date:12/19/2014)... 2014 Research and Markets ( ... "Technology Innovations in Smart Fabrics (Technical Insights)" ... http://photos.prnewswire.com/prnh/20130307/600769 The Research ... a detailed assessment on technological advancements and market ... 1. The Smart Fabrics market is ...
(Date:12/19/2014)... Reports from CDC show approximately ... one or more chronic health conditions - including cardiovascular ... some of these diseases, but may not eliminate the ... While implanted nerve stimulation devices today offer relief, they ... can induce side effects. , To eliminate the ...
(Date:12/19/2014)... Mass. , Dec. 18, 2014  Decision Resources ... dental biomaterial market will grow moderately through 2023 ... procedures and growing awareness of dental biomaterials all spur ... markets such as China and ... will bring dental procedures within reach for a larger ...
Breaking Biology Technology:BioPlus Specialty Pharmacy Promotes Dr. Nick Maroulis to Vice President of Specialty Pharmacy Services 2Technology Innovations in Smart Fabrics (Technical Insights) 2Draper Wins GSK Funding for Smaller, Smarter Implantable Devices 2Asia Pacific Dental Biomaterial Market Will Grow Strongly Through 2023 2Asia Pacific Dental Biomaterial Market Will Grow Strongly Through 2023 3
... Development of Nano-Materials for Angiogenesis Applications , ... OmegaGenesis announced today that they have entered into a ... nano-materials used for Angiogenesis applications. , , ... variety of medical applications ranging from tissue reactivation and healing ...
... 3 Combination Rule Trial for NDA Submission , ... N.J., Dec. 15 QRxPharma Limited (ASX: QRX and ... the development and commercialization of therapies for pain and ... a comparative study to evaluate the efficacy and safety ...
... N.Y., Dec. 15 Allscripts,announced today that ... multispecialty group practice, has selected Allscripts,Practice Management ... for their 190 physicians. , ... ) , ...
Cached Biology Technology:OmegaGenesis Announces Angiogenesis Application Collaboration 2OmegaGenesis Announces Angiogenesis Application Collaboration 3QRxPharma Initiates Comparative Study for MoxDuoIR(TM) Dual-Opioid(TM) Pain Therapy 2QRxPharma Initiates Comparative Study for MoxDuoIR(TM) Dual-Opioid(TM) Pain Therapy 3Community Care Physicians Selects Allscripts Practice Management System for 190 Physicians in New York Capitol Region 2Community Care Physicians Selects Allscripts Practice Management System for 190 Physicians in New York Capitol Region 3Community Care Physicians Selects Allscripts Practice Management System for 190 Physicians in New York Capitol Region 4Community Care Physicians Selects Allscripts Practice Management System for 190 Physicians in New York Capitol Region 5
(Date:12/10/2014)... You,ve been here before: you desperately need to sign into ... key or the answer to your secret question. What,s your ... Today, Hoyos Labs , a digital infrastructure security ... an end to the frustration that comes with usernames, passwords ... a user,s smartphone to acquire his or her biometrics to ...
(Date:12/10/2014)... 08, 2014 Research and Markets ... the "Biometrics Market in Japan 2014-2018" ... http://photos.prnewswire.com/prnh/20130307/600769 The integration of ... rural banking and upgradation of the driver,s license ... the market. Besides the aforementioned projects, biometrics is ...
(Date:12/4/2014)... GARDENS, Fla. , Dec. 3, 2014 /PRNewswire/ ... solutions, today announced that its DigitalPersona ® ... fingerprint readers have been deployed throughout First Bank ... , South Carolina and ... in Southern Pines, North Carolina ...
Breaking Biology News(10 mins):The Password is Finally Dead: Launch of 1U Mobile App Eliminates Need for All Usernames and Passwords 2The Password is Finally Dead: Launch of 1U Mobile App Eliminates Need for All Usernames and Passwords 3Biometrics Market in Japan 2014-2018: Key Vendors are DDS, Fujitsu, Hitachi and NEC 2First Bank Partners with Crossmatch to Increase Log In Security 2First Bank Partners with Crossmatch to Increase Log In Security 3
... survival strategy of the parasite that causes sleeping sickness could ... found that the parasite, which can transform itself into either ... these. One of these types ensures infection in the bloodstream ... by the tsetse fly and spread to another person or ...
... is available in German . It would ... would have similar genetic blueprints. However, scientists from the Max ... with an international research team have now decoded, for the ... ( Arabidopsis lyrata ), a close relative of the thale ...
... release is available in Spanish . ... Agriculture (USDA) scientists and colleagues can improve polymerase chain ... PCR-based tests are prized tools for diagnosing plant ... among other economic harm. But the test,s ability to ...
Cached Biology News:Evolution can cause a rapid reduction in genome size 2Evolution can cause a rapid reduction in genome size 3New technique improves sensitivity of PCR pathogen detection 2
Anti-DDDDK-Tag (Enterokinase Cleavage Site), Agarose Conjugate Research Focus: tag Storage: 4C Shipping Temperature: 4C...
Goat polyclonal to XAGE1 ( Abpromise for all tested applications). Antigen: Synthetic peptide: CGFGFRRQGEDNT, corresponding to C terminal amino acids 149-160 of Human XAGE1 Entrez Gene ID: 9...
Sheep polyclonal to Staphylococcus DNase ( Abpromise for all tested applications). Antigen: Full length native protein (purified)....
Mouse monoclonal [MN-8C4] to zona radiata ( Abpromise for all tested applications). Antigen: Purified eggshell proteins (zona radiata proteins and small amounts of zona pellucida proteins) from ...
Biology Products: