Navigation Links
Quantitative Analysis of GFP in Plant Extracts Using the VersaFluor Fluorometer System

Adrian R. Elliott, Peer M. Schenk and Christopher P.L. Grof
CSIRO Tropical Agriculture, Brisbane, Queensland


Introduction
The Green Fluorescent Protein (GFP) from Aequorea victoria is a useful reporter gene in higher organisms, including plants and animals. The fluorescence emitted is easy to detect and requires no substrate or cofactors. Furthermore, improved GFP versions have been created with higher fluorescence coefficients, improved expression characteristics for eukaryotes, or altered spectral properties. One form, GFP-S65T incorporates a threonine amino acid substitution in place of serine at position 65, which results in red-shifted fluorescence characteristics.1 This form is one that has been proven to be very beneficial for use in plants.

Quantification of GFP is unlike widely used enzyme-based reporters such as β-glucuronidase (GUS) and firefly luciferase (LUC). We show here that fluorescence detection using the VersaFluor fluorometer system with appropriate narrow band filters can be used to detect and quantify GFP-S65T protein levels in plant extracts against a standard curve made from purified GFP-S65T protein.


Method
The test samples consisted of sugarcane cv Q117 callus expressing Ubi1-sgfpS65T-nos3. Approximately 500 mg of tissue was ground in extraction buffer (10 mM Tris-EDTA, pH 8.0, 0.02% [w/v] sodium azide) at 4 C. This extract was centrifuged in a microcentrifuge at 13,000 rpm for 20 min. and the supernatant removed for analysis. Fluorescence microscopy showed that all visible GFP fluorescence was located in the soluble fraction. Protein concentrations of the plant extracts were deter
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Mx4000 Multiplex Quantitative PCR System*,PPP
2. Improved Quantitative Selectivity of Clenbuterol in Human Urine using High Resolution on the TSQ Quantum Mass Spectrometer
3. The LightCycler is the only System for Quantitative PCR that is as Flexible as You are!
4. Quantitative Measurement of Cell Proliferation Using the BrdU ELISA: A Comparison Between Colorimetric and Chemiluminescent Detection
5. Quantitative end-point RT-PCR gene expression measurement using the Agilent 2100 Bioanalyzer and standardized RT-PCR
6. Detection of Genetically Modified Soybean in Processed Foods Using Real-Time Quantitative PCR with SYBR Green I Dye on the DNA Engine Opticon 2 System
7. Qualitative and Quantitative Assessment of Plasmid DNA Isolation, Rev A
8. Quantitative Analysis of Protein-DNA Associations in Vivo Using Real-Time PCR, Rev A
9. Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A
10. Multiplexed Quantitative Peptide Assays for Protein Biomarkers of Cardiovascular Disease in Human Plasma
11. Quantitative Analysis of the Cell Cycle in Imaging Applications
Post Your Comments:
(Date:8/26/2015)... N.J. , Aug. 26, 2015  Roka Bioscience, ... focused on providing advanced testing solutions for the detection ... CEO will present at the Sidoti & Company Emerging ... conference will be held at the New York Marriott ... presentation will be available through the investor relations section ...
(Date:8/26/2015)... FRANCISCO , Aug. 26, 2015  Platform ... that it has received a $1.5M Phase II ... (NIH) to further develop its therapeutic agent to ... clinical need in end stage renal disease (ESRD) ... the National Institute of Diabetes and Digestive and ...
(Date:8/26/2015)... HONG KONG , Aug. 26, 2015 ... "Company"), China,s leading provider of ... and stem cell storage services, today announced that the ... Holdings Limited ("MO2"), an entity wholly owned by Mr. ... the Company, has acquired the outstanding shares (the "Shares") ...
(Date:8/26/2015)... ... August 26, 2015 , ... PRC Clinical, a Clinical Trial ... with local sponsors in southern California. The Clinical Trial Management Expert CRO seeks to ... regulatory pathways., , Stem Cell Meeting on the Mesa ...
Breaking Biology Technology:Symic Biomedical Awarded $1.5MM NIH Phase II SBIR Grant to Develop its Proteoglycan Mimetic Therapeutics to Reduce Vascular Access Failures in ESRD 2Symic Biomedical Awarded $1.5MM NIH Phase II SBIR Grant to Develop its Proteoglycan Mimetic Therapeutics to Reduce Vascular Access Failures in ESRD 3China Cord Blood Corporation Receives Notice of Transactions Regarding its 7% Senior Convertible Notes Due 2017 2China Cord Blood Corporation Receives Notice of Transactions Regarding its 7% Senior Convertible Notes Due 2017 3PRC Clinical To Attend Major Industry Conferences And Meet Sponsor Companies In The San Diego Biotech Hub 2PRC Clinical To Attend Major Industry Conferences And Meet Sponsor Companies In The San Diego Biotech Hub 3
... 26 in Kansas City. , ... Rockville, MD (PRWEB) ... Scientific Meeting (ASM) will be held in Kansas City, Mo., September ... the USP experts responsible for establishing internationally recognized standards for prescription ...
... Oncology Specialty, LP, the,state-of-the-art pharmaceutical distribution center of ... from the National,Safety Council for its outstanding safety ... who produce the "best of the best",in safety ... report,days away from work resulting from workplace injuries ...
... 3 Inverness Medical,Innovations, Inc. (Amex: IMA ... products for the consumer and professional markets, today,announced ... and Jon Russell, Vice President, Finance, are scheduled ... Conference on,Tuesday, September 9th, 2008 in New York. ...
Cached Biology Technology:USP's 2008 Annual Scientific Meeting to Convene Scientific Experts from Around the World 2USP's 2008 Annual Scientific Meeting to Convene Scientific Experts from Around the World 3US Oncology Pharmaceutical Distribution Service Wins National Safety Council Award 2
(Date:8/18/2015)... , Aug. 18, 2015  Navitas today ... contributor to the Submission Content Management webinar ... a non-profit forum. Dedicated to the Implementation of ... neutral platform for industry, vendors, health authorities, ... and work towards a standard method of ...
(Date:8/17/2015)... 17, 2015  NXT-ID, Inc. (NASDAQ: NXTD ) ... on the growing mobile commerce market and creator of ... for Q4 2015. The new marketing campaign is being ... Amplitude Marketing Group ( http://amplitudemarketing.com/ ) and TPMG ( ... leader in retail driven marketing and brand awareness and ...
(Date:8/12/2015)... August 12, 2015 As the ... payment innovation and advanced biometrics technology is accelerating beyond ... consumers rely on using their credit cards, the digital ... future for payment services led by companies in the ... :  Google, Inc. (NASDAQ: GOOG ), Apple Inc. ...
Breaking Biology News(10 mins):Navitas, a Key Contributor in IRISS Forum Webinar 2Navitas, a Key Contributor in IRISS Forum Webinar 3NXT-ID, Inc. Launches Sales Campaign for Wocket Smart Wallet with Award Winning Marketing Team 2NXT-ID, Inc. Launches Sales Campaign for Wocket Smart Wallet with Award Winning Marketing Team 3Digital Wallet Devices & Mobile Payment Management Services Gain a Distinct Advantage in the Digital Payment Evolution as Demand Continues to Grow 2Digital Wallet Devices & Mobile Payment Management Services Gain a Distinct Advantage in the Digital Payment Evolution as Demand Continues to Grow 3Digital Wallet Devices & Mobile Payment Management Services Gain a Distinct Advantage in the Digital Payment Evolution as Demand Continues to Grow 4Digital Wallet Devices & Mobile Payment Management Services Gain a Distinct Advantage in the Digital Payment Evolution as Demand Continues to Grow 5Digital Wallet Devices & Mobile Payment Management Services Gain a Distinct Advantage in the Digital Payment Evolution as Demand Continues to Grow 6
... the traditional summer treat associated with Wimbledon could ... Scientists at the University of Warwick have been ... cardiovascular health, particularly around how they prevent the ... Paul Thornalley from Warwick Medical School heads the ...
... and significance of the biological clocks in polar planktonic ... entitled PolarTime starting July 1st, 2012. It is one ... Association. PolarTime, coordinated by the Alfred Wegener Institute for ... to five years with approximately three million euros from ...
... been made in most developing countries to meet the ... who suffer from hunger by 2015 compared with 1990 ... children,s height and weight, published today in the ... five has improved overall since 1985, one in five ...
Cached Biology News:Just a question of time? 2Just a question of time? 3Developing world has less than 5 percent chance of meeting UN child hunger target, study estimates 2Developing world has less than 5 percent chance of meeting UN child hunger target, study estimates 3
...
Hygromycin resistance gene (hph), provided in a pORF plasmid. pORF is an expression plasmid selectable with Ampicillin....
... 30 isolations. The kit provides a ... apoptotic cells from normal c lture using ... is a Ca2+-dependent phospholipid binding protein with ... Cell Isolation Kit provides a simple and ...
... Linkage: This CLS number is ... to easily match Cornings product ... yet, please order under the ... contact customer service for assistance. ...
Biology Products: