Navigation Links
Optimizing Transfection Conditions for Studying Signal Transduction Pathways

xceeded the level able to be phosphorylated by a limited amount of upstream activators in the cells. Phosphorylated and nonphosphorylated species of the GAL4-ATF2 fusion compete for the GAL4 binding sites on the reporter plasmid. Because transcription of the luciferase gene on the reporter plasmid is activated only by the phosphorylated GAL4-ATF2 fusion protein, lower levels of luciferase activity may occur.

As a negative control, we varied the amounts of the pFA2-dbd plasmidwhich does not contain an activation sequencein parallel transfections in place of the pFA-ATF2 trans-activator plasmid. As expected, this control showed very low background luciferase expression.

Transfecting varying amounts of the expression plasmid containing the gene of interest also had dramatic effects. Hence, another way to optimize induction of the PathDetect system is by transfecting varying amounts of the expression plasmid while maintaining constant amounts of the trans-activator plasmid and reporter plasmid.

In Figure 1, panel B, HeLa cells (1.5 x 105/well) were cotransfected with 10 ng of the pFA-ATF2 plasmid, 0.5 g of the pFR-Luc plasmid, and varying amounts of the pFC-MEKK plasmid. Maximum activation of luciferase expression was seen when 10 ng of the pFC-MEKK plasmid was used per transfection. In this experiment, when the amount of the pFC-MEKK plasmid increased above 10 ng, luciferase activity declined. As before, the GAL4-ATF2 fusion protein is phosphorylated by overexpressing the MEKK protein and binds to the GAL4 binding domain. However, in this scenario, too much of the MEKK protein could cause other signal transduction pathways to overactivate, which would deplete some of the common factors (e.g., coactivators) in the cells transcription machinery. When factors are depleted, they
'"/>

Source:


Page: All 1 2 3 4 5 6 7

Related biology technology :

1. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
2. Optimizing electroporation parameters
3. Optimizing Büchi® Rotary Evaporator Applications
4. Optimizing electroporation parameters
5. Optimizing Büchi® Rotary Evaporator Applications
6. Optimizing Bchi Rotary Evaporator Applications
7. Optimizing electroporation parameters
8. Optimizing electroporation parameters
9. Optimizing electroporation parameters
10. Optimizing electroporation parameters
11. Optimizing electroporation parameters
Post Your Comments:
(Date:11/24/2014)... Continental Clinical Solutions, LLC (Continental) focuses ... at its mid-Atlantic late-phase research facility, located in ... the idea of closing patient disparities in mind. ... advances," said Continental's CEO, Saleh Stevens . ... cell line of Henrietta Lacks, dubious ethical practices ...
(Date:11/23/2014)... November 23, 2014 Apiscent Labs, ... of fine chemical ingredients to the global pharmaceutical ... of its new website, Apiscent.com. , The Apiscent.com ... Labs, clean design, easy navigation, rotating images and ... Labs stand out. , The company’s ...
(Date:11/22/2014)... 2014 RURO, Inc., a leading ... released Limfinity® version 5.1.4. , RURO’s deployment teams ... than ever in RURO’s 8 year history. Many ... enhancement. Limfinity® version 5.1.4 is the perfect example ... bug fixes! , Limfinity® 5.1.4 release highlights – ...
(Date:11/22/2014)...   , ... zu KLOX    Mariano ... Officer bestellt    KLOX ist bestens ... zugelassenen Behandlungssystems zur Wundheilung zu beginnen    , ... sehr erfreut, die folgenden personellen Veränderungen anzukündigen: ...
Breaking Biology Technology:At Continental Clinical Solutions, CEO Saleh Stevens Focuses on Diversity in Clinical Trial Recruitment at Maryland Site 2At Continental Clinical Solutions, CEO Saleh Stevens Focuses on Diversity in Clinical Trial Recruitment at Maryland Site 3RURO Announces Limfinity® version 5.1.4 2KLOX Technologies meldet Neubesetzung von Führungspositionen 2KLOX Technologies meldet Neubesetzung von Führungspositionen 3KLOX Technologies meldet Neubesetzung von Führungspositionen 4KLOX Technologies meldet Neubesetzung von Führungspositionen 5
... against osteoporosis-related fractures, including spine and hip, - Reclast reduced ... to placebo in study published in New England Journal of Medicine, - Unique ... ... - Osteoporotic fractures affect one in two women over 50(1) and are ...
... a biotechnology company developing pharmaceutical products for use ... financing deal. The majority of the funding, $2.3 ... remainder was borrowed from the Madison Development Corporation, ... a private investment firm. The funds raised will ...
... the next strange idea you hear strange: the person ... door. , ,A panel of experienced entrepreneurs speaking ... Milwaukee last Wednesday have dealt with looks and whispers ... in the audience to accept the whispers as a ...
Cached Biology Technology:Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 2Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 3Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 4Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 5Reclast(R) Receives US FDA Approval as First and Only Once-Yearly Treatment for Women With Postmenopausal Osteoporosis 6Quintessence Secures Financing 2Quintessence Secures Financing 3Lowering the fear factor for entrepreneurs, intra-preneurs 2Lowering the fear factor for entrepreneurs, intra-preneurs 3
(Date:11/4/2014)... Nov. 5, 2014 – University of Utah engineers developed ... help jet fuel produce electricity without needing to ignite ... to power portable electronics, off-grid power and sensors. , ... in the American Chemical Society journal ACS Catalysis ... a chemical reaction between a fuel and an oxygen-rich ...
(Date:11/4/2014)...   3D scanner ... Fuel3D , a developer of 3D scanning solutions, ... $6.4 million (£4 million). This funding builds on the company,s ... and paves the way for the commercial launch of the ... round was led by Chimera Partners and will be used ...
(Date:11/3/2014)... 3, 2014  NeuroSigma ® , Inc. (NeuroSigma), ... company focused on commercialization of its non-invasive ... treatment of neurological and neuropsychiatric disorders, announced today ... issued a Notice of Allowance for Mexican Patent ... by NeuroSigma and the Regents of the University ...
Breaking Biology News(10 mins):Jet-fueled electricity at room temperature 2Fuel3D Secures $6.4 Million in Expansion Funding 2Fuel3D Secures $6.4 Million in Expansion Funding 3NeuroSigma Receives Notice of Allowance from the Mexican Institute of Industrial Property Covering Subcutaneous Trigeminal Nerve Stimulation (sTNS) 2NeuroSigma Receives Notice of Allowance from the Mexican Institute of Industrial Property Covering Subcutaneous Trigeminal Nerve Stimulation (sTNS) 3
... Education Association (SEA) is preparing to conduct the first-ever research ... debris in the North Atlantic Ocean. The ... 20-year data set previously collected by SEA that reveals a ... in the western North Atlantic Ocean. This trip ...
... from the University of Warwick has discovered taking too much ... your cholesterol by almost 10%. Selenium is a trace ... from foods such as vegetables, meat and seafood. However, when ... selenium, such as through taking selenium supplements, it can have ...
... , This release is available in French ... more than 3 times per week show a worse performance in ... of exposure to mercury show a generalised delay in cognitive, memory ... fish and canned fish and to a lesser extent in white ...
Cached Biology News:SEA to conduct expedition dedicated to measuring plastic marine debris in the North Atlantic Ocean 2Too much selenium can increase your cholesterol 2Consumption of certain fish during pregnancy associated with poorer cognitive performance 2
... Error-free cloning of BAC (100+kb), fosmid (40kb), ... insert stability than any othe BAC vector. ... high yields and easy recombinant DNA purification. ... transcription-free (pSMART VC) vectors for the highest ...
...
... Adapted to Sf-900 ... were isolated from Spodoptera ... insect cells (1). The ... to serum-free suspension culture ...
6-carboxy-2',4,4',5',7,7'-hexachlorofluorescein, succinimidyl ester (6-HEX, SE) *single isomer*...
Biology Products: