Navigation Links
Ready for Real-time in 5 Minutes

methods to determine gene function. Scientists at Ambion performed a study using cultured HeLa and MCF-7 cells transfected with 3 nM of chemically synthesized siRNA targeting GAPDH to demonstrate the effectiveness of the Cells-to-Signal Kit for measuring knockdown of gene expression (Figure 3). As expected, the expression levels of both GAPDH was down-regulated by more than 80% compared to cells transfected with a scrambled control, while the expression of 18S rRNA was unchanged.

Figure 3. Cells-to-Signal is Ideal for Measuring siRNA Induced Knockdown of Gene Expression. HeLa cells were plated in a 24 well plate at 30,000 cells/well. Cells were transfected with either a scrambled control (green) or with an siRNA to GAPDH (blue) at a concentration of 30 nM. After 48 hours, the cells were harvested and lysed in the Cells-to-Signal Lysis Buffer in a final volume of 500 l. 3 l of the cell lysate was used in one step RT-PCR, using a TaqMan primer and probe set for GAPDH. GAPDH signals were normalized using 18S as an internal control (pink).


Ideal for High Throughput
The quick and simple Cells-to-Signal protocol can be easily adapted to a 96 or 384 well format, thus enabling the simultaneous analyses of multiple cell samples. The protocol will afford excellent reproducibility as seen in Figure 4 where VEG-F and GAPDH expression levels were determined in HeLa and MCF-7 cells grown in a 96 well plate.

Figure 4. Measurement of GAPDH and VEG-F Expression Levels in a 96 well Format Using Cells-to-Signal. 3000 HeLa or MCF-7 cells were plated in 48 wells each o
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood
2. Advantages of ReadyAgarose Precast Gels Over 96-Well Handcast Gels for High-Throughput Analysis, Rev A
3. Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets.
4. Measuring RNase Activity A Real-time Kinetic Analysis
5. YOUR DATA: Direct Real-time RT-PCR on Sympathetic Neuron Lysates Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs
6. YOUR DATA: Direct Real-time RT-PCR on Sympathetic Neuron Lysates Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Ready for Real time Minutes

(Date:8/20/2014)... your ear could improve the health of your heart, ... of Leeds used a standard TENS machine like those ... to the tragus, the small raised flap at the ... ear canal. , The stimulation changed the influence of ... nervous signals that can drive failing hearts too hard. ...
(Date:8/19/2014)... One of the major limitations ... inadequate vascular supply. Nutrient and gas transport throughout ... critical for successful regeneration and integration into the ... in vascularization and promote angiogenesis in a variety ... design and architecture, and enhancing scaffolds with stem ...
(Date:8/19/2014)... A Geisinger researcher on a personal crusade ... for Disease Control and Prevention (CDC) award recognizing his ... Joseph Boscarino , Ph.D. MPH, senior ... of researchers nominated for the Centers for Disease Control ... Data Methods and Study Design. "Hepatitis C ...
(Date:8/19/2014)... 19, 2014 Robin Williams’ passing is ... take on an individual. Symptoms range from slowness ... problems swallowing and severe depression. Parkinson’s disease progressively ... of dopamine producing neurons of the brain. As ... that slowly and progressively gets worse, they look for ...
Breaking Biology Technology:'Tickling' your ear could be good for your heart 2Vascularization in Tissue Engineering Highlighted at 12th New Jersey Symposium on Biomaterials Science 2Geisinger investigator nominated for federal award 2Geisinger investigator nominated for federal award 3As We Mourn Robin Williams’ Passing, His Death Sheds Light on Patients Struggling with Parkinson’s Disease 2As We Mourn Robin Williams’ Passing, His Death Sheds Light on Patients Struggling with Parkinson’s Disease 3
... Burn rate significantly reduced and Semen Sexing Technology achieves, ... /PRNewswire-FirstCall/ - For nine months ended June 30, 2009, Microbix ... burn rate. The third quarter had a small sales retraction ... rates. The forecast is for continued growth in sales. , ...
... , , NEWTON, Mass., Aug. 6 ... the Company will release second quarter 2009 financial results on ... markets. , , Michael C. Ferrara, President and ... and Chief Accounting Officer, will host a conference call and live ...
... , PALO ALTO, Calif., Aug. 6 Telik, ... $5.7 million, or $0.11 per share, for the second quarter ended June ... per share, for the comparable period in 2008. , , ... expenses were $6.3 million, compared with $11.0 million in the 2008 second ...
Cached Biology Technology:Microbix Sales Grow 22% For Nine Months 2Microbix Sales Grow 22% For Nine Months 3Microbix Sales Grow 22% For Nine Months 4Microbix Sales Grow 22% For Nine Months 5Microfluidics International Corporation to Report Second Quarter 2009 Financial Results on August 11, 2009 2Telik Announces Second Quarter 2009 Financial Results 2Telik Announces Second Quarter 2009 Financial Results 3Telik Announces Second Quarter 2009 Financial Results 4
(Date:8/20/2014)... When a colony of honeybees grows to about 4,000 ... reproductive cycle: the building of a special type of ... team of experts from the Department of Neurobiology and ... what starts the reproductive cycle of honeybee colonies. The ... The Science of Nature . , Reproduction isn,t ...
(Date:8/19/2014)... be made to decrease China,s increasing caesarean section rate, ... BJOG: An International Journal of Obstetrics and Gynaecology ... caesarean delivery rates in the world. Of 16 million ... Although the exact rate is not known, the current ... total caesarean rates ranging from 36% to 58%. However, ...
(Date:8/19/2014)... lemur. It,s not an African Bush Baby or even a ... downright "cool" primate from Southeast Asia. , "It,s really not ... Brown, curator-in-charge at the University of Kansas, Biodiversity Institute. "A ... body; a long tail with a furry tuft at the ... a bit like the disks on the digits of tree ...
Breaking Biology News(10 mins):Worker bees 'know' when to invest in their reproductive future 2Is China's 50 percent cesarean section delivery rate too high? 2Is China's 50 percent cesarean section delivery rate too high? 3Philippine tarsier gets boost from Kansas research, and genetic proof of a new variety 2Philippine tarsier gets boost from Kansas research, and genetic proof of a new variety 3
... DENVER Researchers have linked a variant in the ... chronic obstructive pulmonary disease (COPD) in Caucasian men. The ... Normative Aging Study, a multidisciplinary study of aging that ... presented at the ATS 2011 International Conference. "Our ...
... the process of converting the genetic information encoded in ... protein or any of several types of RNA. Scientists ... different physiological processes throughout the body are robustly pre-determined ... new study by researchers from the University of California, ...
... Most people who have had the experience of having ... the animals can "recognize" us. They seem to recognize our ... they respond to us differently from other people. , Actually, ... shown that domesticated animals, such as honey bees, chickens, pigeons, ...
Cached Biology News:Gene variant linked with development of COPD in men 2Plasticity of hormonal response permits rapid gene expression reprogramming 2I know you, bad guy! 2I know you, bad guy! 3
The Discovery Series PDQuest 2-D analysis software is used for imaging, analyzing, and databasing two-dimensional electrophoretic gels. The 10-user network license entitles ten users to access data f...
Mouse monoclonal antibody raised against a partial recombinant NR4A2. NCBI Entrez Gene ID = NR4A2...
... Polymorphic DNA Technologies also offers a DNA ... of templates including plasmids, PCR products, BACs, ... customer-provided templates such as purified plasmids or ... liquid handling robotics optimized for automated reaction ...
... NightOWL is a dedicated imaging system ... size. Equipped with a very sensitive camera ... or plants in tumor research, infectious disease ... Western blots. NightOWL is the only imager ...
Biology Products: