Navigation Links
Real-Time Immuno-PCR on the iCycler iQ System, Rev A

Kristina Lind and Mikael Kubista, TATAA Biocenter, Medicinaregatan 7A/B, 405 30 Gteborg, Sweden


Introduction
A technique for antigen detection, called immuno-PCR, was developed by Sano et al. (1992). It combines the molecular recognition of antibodies with the high DNA amplification capability of PCR. The procedure is similar to conventional enzyme-linked immunosorbent assays (ELISA) but allows for more sensitive detection. Instead of an enzyme, a DNA molecule is linked to the detection antibody and serves as a template for PCR (Figure 1). The DNA molecule is amplified and the PCR product is measured by gel electrophoresis. An improvement of this method is to amplify the oligomer in a real-time PCR instrument, thereby eliminating post-PCR analysis (Sims et al. 2000). Further, real-time PCR is extremely accurate and sensitive, which should make it possible to quantitate very low amounts of DNA-coupled detection antibody with high accuracy. Here we present early results on the development of real-time immuno-PCR for prostate specific antigen (PSA) using the iCycler iQ system. PSA is a well-known tumor marker for prostate cancer and is widely used to detect, stage, and monitor the disease.


Methods
Anti-PSA10 and anti-PSA66 from CanAg Diagnostics were used as capture and detection antibodies, respectively, in the sandwich immuno-PCR assay. Anti-PSA66 was biotinylated using biotinamido-caproate-N-hydroxysuccinimide ester. Biotinylated DNA was generated by amplifying a 1,098 bp fragment of the gusA gene (E. coli β-glucuronidase gene) with a 5'-biotinylated forward primer (biotin-AACTATGCCGGAATCCATCG- 3') and unmodified reverse p
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. 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
2. Monitoring Microbial Populations Using Real-Time qPCR on the MJ Research Opticon 2 System
3. Rapid SNP Genotyping of the Pharmacogenomically Important Allele CYP2D6*4 Using Real-Time qPCR on the DNA Engine Opticon 2 System
4. Real-Time Multiplex PCR from Genomic DNA Using the iCycler iQ Detection System
5. Quantitative Analysis of Protein-DNA Associations in Vivo Using Real-Time PCR, Rev A
6. Real-Time Quantification of Genomic DNA Using DyNAzyme II DNA Polymerase and SYBR Green I Dye
7. Quantitation of Lymphangiogenesis Using the iCycler iQ Real-Time PCR Detection System and Scorpions Detection System, Rev A
8. Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A
9. Real-Time PCR/Melt-Curve Analysis: SNP Detection With FRET, Rev A
10. Real-Time PCR: General Considerations, Rev A
11. Identification of Nonspecific Products Using Melt-Curve Analysis on the iCycler iQ Detection System, Rev A
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Real Time Immuno PCR the iCycler System Rev

(Date:10/22/2014)... The Americas Inorganic Refrigerants Market report ... Americas with analysis and forecast of revenue. , ... Refrigerants Market report, to get an idea of ... glimpse of the segmentation in the Americas inorganic ... and figures. , http://www.micromarketmonitor.com/market/americas-inorganic-refrigerants-4191654241.html , ...
(Date:10/22/2014)... October 22, 2014 Currently offering ... A & D Weighing, Rice Lake, and Ohaus, ... Balance . The Sartorius CPA Semi-Micro Balance is ... high-quality, precise, and user-friendly laboratory balance. , Sartorius ... equipment, their laboratory balances are well known for ...
(Date:10/22/2014)... VANCOUVER , Oct. 22, 2014 /PRNewswire/ - iCo ... ICOTF), today announced next steps for its Oral ... findings from its in vitro work ... therapy.  iCo now plans to complete pre clinical ... an initial Phase 1A clinical trial, utilizing approximately ...
(Date:10/20/2014)... 2014 Mapp Biopharmaceutical,s valiant effort to ... to fight the Ebola outbreak will make the ... production of pharmaceuticals can be, according to Kalorama ... while some may be taken aback by the ... industry knowledge are well aware of the complex ...
Breaking Biology Technology:The Americas Inorganic Refrigerants Market is estimated to grow to $71.6 million by 2018 - New Report by MicroMarket Monitor 2The Americas Inorganic Refrigerants Market is estimated to grow to $71.6 million by 2018 - New Report by MicroMarket Monitor 3Pipette.com Launches the Addition of Sartorius CPA Semi-Micro Balance to Their Product Portfolio with Special Promotional Pricing 2Pipette.com Launches the Addition of Sartorius CPA Semi-Micro Balance to Their Product Portfolio with Special Promotional Pricing 3iCo Therapeutics Announces Advancement of Oral Amphotericin B Program 2iCo Therapeutics Announces Advancement of Oral Amphotericin B Program 3Kalorama: ZMapp Highlights Need For Faster Biopharmaceutical Production 2Kalorama: ZMapp Highlights Need For Faster Biopharmaceutical Production 3
... of Madison has been licensed to sell a ... agents, GenTel announced on Tuesday. , ,GenTel will market ... Kreatech Biotechnology BV of Amsterdam, under its ... are validated for use on GenTel's PATH protein microarray ...
... drive thru Illinois you now pay a premium for using cash ... device for your car. If you stop for gas at a ... plastic device on their key chain to pay for gas. These ... applications to reduce costs, improve efficiency, and potentially improve the buyers ...
... Wis. - Former Wisconsin Gov. and Secretary of ... board of directors at C.R. Bard , the ... at the company's August board meeting. , ,Bard, a ... sells specialty devices for vascular, urology, oncology and surgical ...
Cached Biology Technology:
(Date:10/17/2014)... regarding Ebola is critical and pertinent for practicing physicians ... outbreak and epidemic. The Journal, Disaster Medicine and ... Ebola Virus and Public Health, to surround the public, ... critical societal moment. , On October 17, the ... . The primer was prepared by Dr. Eric Toner, ...
(Date:10/16/2014)... wages war on the human body. Battles are ... In pancreatic cancer, this stalemate—known as tumor dormancy—can ... malignant, a phenomena that is poorly understood. ... laboratory of Salvatore Torquato, a Professor of Chemistry ... surrounding tumor dormancy and the switch to a ...
(Date:10/16/2014)... cultivated efficiently, they are anything but sustainable: environmental ... cultivation is becoming increasingly evident. Despite their disadvantages, ... are regarded as the sole possibility of achieving ... finds Bernhard Schmid, an ecology professor at the ... of agriculture and forestry. After all, a new ...
Breaking Biology News(10 mins):Modeling tumor dormancy 2Plant communities produce greater yield than monocultures 2Plant communities produce greater yield than monocultures 3
... is measured to understand tree health, fluxes in carbon ... of the most essential and widely collected woody plant ... is awkward and time consuming. Scientists have developed a ... safely and accurately. Dendrometer bands are metal straps ...
... (IL-7) is an important immune messenger protein which ensures ... in our body for immune defence. Researchers from ETH ... function: it enhances the drainage function of lymphatic vessels, ... vessels into the body tissue and return it to ...
... an old dog new tricks." Now neuroscientists are beginning to ... neuroscientists have struggled to understand how the microcircuitry of the ... for the old. New findings published in the journal ... Los Angeles and the University of California, Irvine show how ...
Cached Biology News:New technique for measuring tree growth cuts down on research time 2New technique for measuring tree growth cuts down on research time 3New function for a well-known immune messenger molecule 2New function for a well-known immune messenger molecule 3Researchers discover how inhibitory neurons behave during critical periods of learning 2
... HISTAR detection systems provide linking and ... with species specific primary antibodies for ... Visualisation is accomplished using an ultra-sensitive ... polymer labelling technology. Secondary antibodies are ...
... Enzyme concentration - 5 units/µL ,• High ... yields ,• Leaves an 'A' overhang ,• ... which exhibits very high activity in primer ... ensure the quality of the enzyme, each ...
... from Thermus aquaticus expressed in E. coli; ... of 250 units enzyme/tube and Buffer I ... the same enzyme as AmpliTaq DNA Polymerase ... separation process to ensure that contaminating bacterial ...
Our original highly-purified, robust Taq Polymerase which delivers consistently high yields and minimal background...
Biology Products: