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:1/14/2014)... York, NY (PRWEB) January 14, 2014 ... of Alliqua, Inc. (OTCQB: ALQA). Alliqua is an emerging ... to serve the wound care market. , Free report ... was restructured with a seasoned management team and Board, ...
(Date:1/14/2014)... In recent years, growing suspicion about the pharmaceutical ... and promotion has led to unprecedented levels of public ... about the insidious impact of commercialization of research, has ... the world’s biggest pharmas for illegal marketing activities, allegations ...
(Date:1/14/2014)... USA, and Cardiff, UK (PRWEB) January 13, 2014 ... technology development leader with more than 20 years in ... for optics and photonics . Hainsey will serve as ... delighted to have Dr. Hainsey join SPIE as our ...
(Date:1/14/2014)... (PRWEB) January 14, 2014 BioMedomics, Inc. ... ultra-sensitive POC diagnostic platforms and novel disease specific POC tests, ... of financing netting a total of $690,000. The investment is ... in the company. This group of private investors has significant ...
Breaking Biology Technology:EquitiesIQ Initiates Coverage of Alliqua, Inc. 2EquitiesIQ Initiates Coverage of Alliqua, Inc. 3The Sunshine Act: Necessary Regulation or Unnecessary Dysregulation? New Life Science Webinar Hosted by Xtalks and IRB Services 2The Sunshine Act: Necessary Regulation or Unnecessary Dysregulation? New Life Science Webinar Hosted by Xtalks and IRB Services 3Photonics R&D Leader Bob Hainsey Joins SPIE Technical Staff 2BioMedomics, Inc. Closes $690K Round of Financing 2
... CARY, N.C., May 19, 2011 ... its flexible 100% web-based Life Science Study Management ... successfully used globally by Leading European and USA ... eStudy,s on-line study design, management, communication, data collection ...
... CLARA, Calif., May 19, 2011 The 2012 federal ... National Nanotechnology Initiative (NNI), a federal interagency research and ... manipulation of matter at the nanoscale—a measurement of particles ... meter) in size.  While nanoscale materials are found in ...
... Inc., a Massachusetts-based biopharmaceutical company, announced today that it ... Administration on a Special Protocol Assessment (SPA) for a ... agent for patients with newly diagnosed prostate cancer. The ... with definitive results expected by 2015. The randomized study ...
Cached Biology Technology:Leading Ag Biotechnology Companies in Europe and America Save 50-80% of Field Study Management Time with eStudy 2Leading Ag Biotechnology Companies in Europe and America Save 50-80% of Field Study Management Time with eStudy 3Environmental Training Center Prepares Companies to Handle Nanotechnology Environmental and Human Safety Impacts 2Advantagene Announces SPA Agreement With FDA to Launch Phase 3 Trial for Novel Vaccine Aimed at Preventing Prostate Cancer Recurrence 2
(Date:4/17/2014)... NJ. April 16, 2014. Kessler Foundation has been ... million from the Department of Defense Spinal Cord ... principal investigator for the randomized, double-blinded, controlled, multi-site ... bone and muscle strength after spinal cord injury. ... & Engineering Research at Kessler Foundation. Two additional ...
(Date:4/17/2014)... births, Down syndrome - or trisomy 21 - is ... results from a chromosomal abnormality where cells of affected ... of the human genome). A study conducted by Stylianos ... Medicine and Development at the University of Geneva (UNIGE) ... light on how the extra chromosome 21 upsets the ...
(Date:4/17/2014)... ago, Katia Silvera , a postdoctoral scholar at the ... field trip in a mountainous area in central Panama when ... , Unable to identify it, they contacted German Carnevali, a ... be an unnamed species. So Carnevali recently named it after ... genus name, comprising about 40 species in the world. ...
Breaking Biology News(10 mins):Kessler Foundation awarded Department of Defense grant for spinal cord injury research 2Trisomy 21: How an extra little chromosome throws the entire genome off balance 2Trisomy 21: How an extra little chromosome throws the entire genome off balance 3Orchid named after UC Riverside researcher 2Orchid named after UC Riverside researcher 3
... fruit flies eat like horses. Hatching inside over-ripe fruit where they ... sends them an offer they cant refuse. To survive, they must ... they avoid food as if it were poison. Only then can ... to lay their own eggs. Now, a team of researchers ...
... migration to EAC ePassport technology, DALLAS, May ... (Nasdaq: ENTU ) will provide its proven ... authenticate sensitive,biometric information stored on machine readable travel ... will be available to nearly 23 million,citizens by ...
... Board: BKYI) today announced plans to release first quarter,2008 financial ... conjunction with the release, BIO-key has scheduled a conference call,which ... 15, 2008 at,9:00 a.m. Eastern Time., What: ... Call ...
Cached Biology News:Fruit fly avoidance mechanism could lead to new ways to control pain in humans 2Entrust, Hewlett Packard Partner to Deploy Taiwanese ePassports, Authenticate Biometric Data 2Entrust, Hewlett Packard Partner to Deploy Taiwanese ePassports, Authenticate Biometric Data 3BIO-key(R) Announces First Quarter 2008 Earnings Release and Conference Call Schedule 2
... Captivate magnetic separator for 96-well ... in conjunction with the Captivate ... or other magnetic particles and ... microplates. Cell Biology Cell Separation ...
RABBIT ANTI RAT AQUAPORIN 5...
MOUSE ANTI S. NEWPORT SALMONELLA HADAR...
Secure-Seal™ hybridization chamber gasket, eight chambers, 9 mm diameter, 0.8 mm deep *set of 20*...
Biology Products: