Navigation Links
Measurement of Green Fluorescent Protein in the SpectraMax Gemini XS Spectrofluorometer (MaxLine Application Note #44)

a laser scanning confocal microscope or similar instrument. A weak signal thus precludes use in a microplate spectrofluorometer, which would allow increased throughput. The SPECTRAmax GEMINI-XS is a dual-monochromator microplate fluorometer system incorporating highly sensitive optics4 that permit experimental determinations at low-level detection limits. This application note describes experimental protocols that enable the detection of relatively low numbers of cells expressing Green Fluorescent Protein (GFP) in a 96-well microplate using the SPECTRAmax GEMINI-XS spectrofluorometer.


MATERIALS AND METHODS
Human embryonic kidney 293 cells (HEK-293) were transiently transfected with a wild-type GFP expression plasmid (pS65T, Clontech, discontinued product; cytomegalovirus immediate early promoter drives expression of GFP) using Lipofectamine as per manufacturer's instructions (Gibco/Invitrogen, Cat. No. 18324-012). Cells were grown to confluence in 162 cm2 flasks under standard cell-culture conditions. Fluorescence microscopy revealed an expression level in excess of 75 %.

The GFP-expressing cells were removed with trypsin, and the cells were washed with medium (MEM Alpha, Gibco-RBL Cat. No. 22571-020). The appropriate number of cells was seeded in alternate columns of a 96-well microplate (Costar, black walled, clear bottomed Cat. No. 3603) and diluted with mock-transfected cells to maintain a constant cell number of 200,000 cells per well. This resulted in a final number of 6,250 (column 12) to 200,000 (column 2) GFP expressing cells per well in a volume of 100 l. These cells were seeded in alternate columns 2, 4, 6, 8, 10, and 12. In contrast, columns 1, 3, 5, 7, 9 and 11 were seeded with 200,000 mock-transfected cells per well. Then the plate was returned to the incubator for 48 hours.

Initially, excitation and emission scans were run to select the optimum excitation wa
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. Quantitative Measurement of Cell Proliferation Using the BrdU ELISA: A Comparison Between Colorimetric and Chemiluminescent Detection
2. Cell Proliferation and Viability Measurement
3. LabVIEW for Measurement and Data Analysis
4. Use of FlashPlate Technology for In Vitro Measurement of [35S]-GTPγS Binding in CHO Cells Expressing the Human 5-HT1B Receptor
5. Use of FlashPlate Technology for In Vitro Measurement of 125I-Labeled TGF-1 Binding on Chimeric Extracellular Domain of Type II Transforming Growth Factor Receptor
6. In Vitro Measurement of the Second Messenger cAMP: RIA vs. FlashPlate
7. Measurement of proteasome inhibition in live cells in Molecular Devices microplate fluorometers
8. Measurement of Molecular Beacons in the SpectraMax Gemini Spectrofluorometer (MaxLine Application Note #36)
9. PathCheck Applied to Measurement of Protein Solutions in the SpectraMax Plus Microplate Spectrophotometer (MaxLine Application Note #26)
10. UV Absorbance Measurements in SpectraMax Microplate Spectrophotometers MaxLine Application Note #32)
11. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
Post Your Comments:
(Date:7/30/2015)... ... , ... The 2015 Market Research Report on the Global Propanol Industry ... Propanol market with a focus on the Chinese situation. Major companies included in the ... Chemical, Sanofi, Royal Dutch Shell, LG Chem and Zhejiang Xinhua Chemical. , The report ...
(Date:7/30/2015)... 2015   Senomyx , Inc. (NASDAQ: SNMX ... to discover, develop, and commercialize novel flavor ingredients for ... financial results for the second quarter 2015. ... remain on track to achieve our commercial and financial ... of the Company. "Since our last quarterly earnings report, ...
(Date:7/30/2015)... Pa. , July 30, 2015  Discovery ... that it has received the second $1.0M tranche ... Innovation Research (SBIR) grant valued at up to ... company,s aerosolized KL4 surfactant as a potential medical ... injury.  Discovery Labs was awarded an initial $1.0 ...
(Date:7/29/2015)... July 30, 2015 Sanofi, ... publie ses résultats pour le premier semestre 2015. ... les résultats. Visionner l,interview vidéo et ... Au sommaire de l,interview :  ... Moteurs de croissance - Diabète ...
Breaking Biology Technology:Global Propanol Market Technical Data and Manufacturing Analysis 2020 2Global Propanol Market Technical Data and Manufacturing Analysis 2020 3SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 2SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 3SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 4SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 5SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 6SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 7SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 8SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 9Discovery Labs Receives $1.0 million under an SBIR Grant in Continuing Support of the Development of Aerosolized KL4 Surfactant to Address Radiation-Induced Lung Injury 2Discovery Labs Receives $1.0 million under an SBIR Grant in Continuing Support of the Development of Aerosolized KL4 Surfactant to Address Radiation-Induced Lung Injury 3
... CALUIRE, France, Feb. 19 Setaram,Inc., a ... announces the acquisition of Hy-Energy LLC (Newark, ... scientific instruments for,the ever-expanding field of clean ... a full suite of analytical capabilities which,complement ...
... 19 Barr Pharmaceuticals,Inc. (NYSE: BRL ) ... received final approval from the U.S. Food and ... Ingelheim,Pharmaceuticals, Inc.,s Mirapex(R) Tablets (Pramipexole Dihydrochloride),0.125mg, 0.25mg, 0.5mg, ... is the,first to file an Abbreviated New Drug ...
... Technology Providers Broadens ... Opportunities for Distribution, New Markets, SILVER SPRING ... and Immersion Medical, a subsidiary of Immersion Corporation,announced ... enhanced version of B-Line Medical,s SimCapture(TM) product. SimCapture,is ...
Cached Biology Technology:Setaram Inc. Acquires Hy-Energy LLC 2Barr Receives Final Approval for Generic Mirapex(R) Tablets, 0.125mg, 0.25mg, 0.5mg, 1mg & 1.5mg 2Barr Receives Final Approval for Generic Mirapex(R) Tablets, 0.125mg, 0.25mg, 0.5mg, 1mg & 1.5mg 3Barr Receives Final Approval for Generic Mirapex(R) Tablets, 0.125mg, 0.25mg, 0.5mg, 1mg & 1.5mg 4B-Line Medical and Immersion Medical Announce Partnership 2B-Line Medical and Immersion Medical Announce Partnership 3
(Date:7/13/2015)... 2015 NXT-ID, Inc. (NASDAQ: NXTD ) ... focused on the growing mobile commerce market, announced ... for BEHAVIORAL-DIRECTED AUTHENTIATION METHOD AND SYSTEM. ... secure method to make payments.  With this patent, ... with its groundbreaking voice-direct payment patent application, with ...
(Date:7/8/2015)... 2015 Summary Pancreatic cancer is the ... fatal, with a mortality rate of 10.9 deaths per ... cancer patients has highlighted a significant need for new ... met by the current market. A highly active ... varying molecule types and mechanisms of action, which provides ...
(Date:7/7/2015)... VIEW, Calif. , July 7, 2015 /PRNewswire/ ... biometrics market, Frost & Sullivan recognizes Credence ID, ... Sullivan Entrepreneurial Company of the Year Award. ... suite to aid in its mission of offering ... under-recognized populations globally. As Credence ID was formed ...
Breaking Biology News(10 mins):NXT-ID Patents Behavior-Directed Payments 2NXT-ID Patents Behavior-Directed Payments 3NXT-ID Patents Behavior-Directed Payments 4Frontier Pharma: Pancreatic Cancer - Identifying and Commercializing First-in-Class Innovation 2Frontier Pharma: Pancreatic Cancer - Identifying and Commercializing First-in-Class Innovation 3Frost & Sullivan Acknowledges the Distinctive Customer-focused Attributes of Credence ID's Mobile Biometric Product Suite 2Frost & Sullivan Acknowledges the Distinctive Customer-focused Attributes of Credence ID's Mobile Biometric Product Suite 3Frost & Sullivan Acknowledges the Distinctive Customer-focused Attributes of Credence ID's Mobile Biometric Product Suite 4Frost & Sullivan Acknowledges the Distinctive Customer-focused Attributes of Credence ID's Mobile Biometric Product Suite 5
... pollution most often produced from biomass burning, cooking with ... in the atmosphere three to four times greater than ... article in the journal Nature Geoscience. Scripps Institution ... Ramanathan and University of Iowa chemical engineer Greg Carmichael, ...
... REST blocks the expression of a microRNA that prevents ... to differentiate into specific cell types, scientists at The ... in the journal Nature. Researchers show RE1-silencing ... stem cells, said senior author Sadhan Majumder, Ph.D., professor ...
... may cluster the genes needed to make defence chemicals. Their ... products of use as drugs, herbicides or crop protectants. Using ... as a template, they uncovered a previously unknown gene cluster ... now want to extend the search to other plants. ...
Cached Biology News:Black carbon pollution emerges as major player in global warming 2Black carbon pollution emerges as major player in global warming 3Protein protects embryonic stem cells' versatility and self-renewal 2Protein protects embryonic stem cells' versatility and self-renewal 3Plant gene clusters for natural products 2
Anti-Human Factor XIII HRPO (Polyclonal) (sheep IgG)...
Recognizes the 41 and 43 kDa G-proteins present in liver plasma membranes and Gq/11alpha expressed in the Sf9 insect cell expression system. Negative for recombinant G(I)alpha subtypes....
Rabbit polyclonal to Flagellin ( Abpromise for all tested applications). Antigen: Synthetic peptide from the flagellin C terminal domain of Burkholderia pseudomallei....
This application changes sensitive keywords in an FCS file to remove patient information. It was contributed by Dr. David Miller....
Biology Products: