Navigation Links
Setting up a kinase profiler with IMAP

L with specific activity of 1000 units per mg provides 40 units/mL, where 1 unit is defined as 1 nmol phosphate incorporated into substrate per minute.

Step 2. An EC50 concentration (predetermined by enzyme titration assay) of each kinase is suggested for the screen set up in Table B. Prepare a working stock of 150 mL of 4X the EC50 concentration for each kinase on one plate.


Prepare the ATP Working Solutions
Step 1. To make an 800 M ATP working solution, add 0.16 mL of a 10 M stock of ATP per 1.84 mL of Complete Reaction Buffer.

Step 2. To make an 80 M ATP working solution, add 0.1 mL of the 800 M solution prepared in Step 4A to 0.9 mL of Complete Reaction Buffer.

Step 3. The resulting dilutions of 800 and 80 M are 8X the final reaction concentrations of 100 and 10 M ATP.


Prepare the Substrate Working Solutions
For this assay, there are 10 different substrate solutions to prepare. Refer to Table A for details.

Step 1. Prepare each 20 M Fluorescent Substrate solution in Reaction Buffer with no DTS. Aliquot and store substrates at -20C to asssure stability for future use.

Step 2. To make an 800 nM substrate working solution, add 20 L of each 20 M solution per 480 L of Complete Reaction Buffer. Aliquot 3 x 150 L per microtube. This solution is 8X the final reaction concentration of 100 nM substrate.


B: SET UP KINASE REACTION
Add components to the 384-well assay plate according to the plate diagram in Table B

'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

Related biology technology :

1. Setting up the laboratory to avoid contamination
2. Reduce Errors and Save Time When Setting Up PCRs
3. Setting up Successful siRNA Library Screens
4. LiquiChip Kits for bead-based cytokine and kinase assays
5. Homogeneous, chemiluminescent kinase assay kits for assessing the functional activity of a broad range of serine-threonine and tyrosine kinases
6. TKXtra DataFile: Homogenous tyrosine kinase detection using High Efficiency Fluorescence Polarization
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Setting kinase profiler with IMAP

(Date:1/15/2014)... NC (PRWEB) January 15, 2014 DTS ... to its Online Web Portal for Life Science organizations who ... to specify the subject matter of their documents in advance ... will help reduce time-to-delivery of translations, often a critical factor ...
(Date:1/14/2014)... Doylestown, PA (PRWEB) January 14, 2014 Date: ... p.m. , Location: Warrington Country Club, 1360 Almshouse Road, Warrington, ... only national nonprofit organization solely dedicated to finding a cure ... those affected worldwide, will host its annual Crystal Ball on ...
(Date:1/14/2014)... 14, 2014 EquitiesIQ, a leading informational ... ALQA). Alliqua is an emerging biomedical company acquiring, developing, ... care market. , Free report download: http://equitiesiq.com/reports/alliqua/ ... seasoned management team and Board, which launched the company’s ...
(Date:1/14/2014)... Jan. 14, 2014  RXi Pharmaceuticals Corporation (OTCQX: RXII), ... commercializing innovative therapies addressing major unmet medical needs ... the Notice of Allowance from the United States ... RNAi compounds (sd-rxRNA®), for the treatment of fibrosis. ...
Breaking Biology Technology:DTS Improves Efficiency for Life Science Document Translations 2Hepatitis B Foundation to Host Annual Crystal Ball Gala 2EquitiesIQ Initiates Coverage of Alliqua, Inc. 2EquitiesIQ Initiates Coverage of Alliqua, Inc. 3RXi Pharmaceuticals Receives US Notice of Allowance for a Key Patent Relating to its Self-Delivering Technology with sd-rxRNAs targeting CTGF, including RXI-109, for the Treatment of Fibrotic Disorders 2RXi Pharmaceuticals Receives US Notice of Allowance for a Key Patent Relating to its Self-Delivering Technology with sd-rxRNAs targeting CTGF, including RXI-109, for the Treatment of Fibrotic Disorders 3
... Corporation (NASDAQ: LIFE ) today announced that ... open-access resource for Ion Torrent users and developers, has ... sudden jump in membership began after Ion Torrent published ... Ion Personal Genome Machine™ (PGM™), the world,s first benchtop ...
... 2011 BioNeutral Group, Inc. (OTCBB: BONU), a ... it has received $400,000 in connection with the ... Pty Ltd ("Vinfluence"), New South Wales, Australia.  Pursuant ... from the initial closing and a second funding ...
... computer assisted design (CAD) tools that made it ... transistors may soon be coming to the biological ... (DOE),s Joint BioEnergy Institute (JBEI) have developed CAD-type ... it possible to engineer biological components or "RNA ...
Cached Biology Technology:Number of Ion Community Users Triples, Spurred by Open Access to DNA Sequencing Protocols and Analysis Software 2Number of Ion Community Users Triples, Spurred by Open Access to DNA Sequencing Protocols and Analysis Software 3Number of Ion Community Users Triples, Spurred by Open Access to DNA Sequencing Protocols and Analysis Software 4Number of Ion Community Users Triples, Spurred by Open Access to DNA Sequencing Protocols and Analysis Software 5BioNeutral Group Receives $400,000 in Equity Financing 2BioNeutral Group Receives $400,000 in Equity Financing 3CAD for RNA 2CAD for RNA 3CAD for RNA 4
(Date:4/17/2014)... A Kansas State University engineer has developed a ... detects improvised explosive devices. The same technique could ... the Steven M. and Kay L. Theede chair ... nuclear engineering, and his research team have created ... or in car trunks. The distance detection method ...
(Date:4/17/2014)... The White House honored Clemson professor Rajendra Singh ... efforts to promote and expand solar deployment in ... the D. Houser Banks Professor of Electrical and ... Silicon Nanoelectronics, is considered a local hero leading ... and economic opportunity in solar power and driving ...
(Date:4/17/2014)... deforestation and fragmentation of forests in the Amazon ... forests, contributing to rapid and widespread forest loss ... researchers., The findings show that forests in the ... droughts coupled with forest fires lead to large-scale ... Jennifer Balch, assistant professor of geography, Penn State. ...
Breaking Biology News(10 mins):Patented research remotely detects nitrogen-rich explosives 2White House honors Clemson professor as 'Champion of Change' for solar deployment 2Drought and fire in the Amazon lead to sharp increases in forest tree mortality 2Drought and fire in the Amazon lead to sharp increases in forest tree mortality 3
... and Imperial College London today signed an official agreement ... was previously announced by Prime Minister Lee Hsien Loong ... the working name for the school is "Imperial College ... start in 2013, ICNMS will be positioned to meet ...
... Department of Energy,s Joint BioEnergy Institute (JBEI) have ... the development of a next generation of clean, ... economics and well as performance. This on-line, wiki-based ... strategies for cost-efficient biorefinery operations by simulating such ...
... Scientists of the Max Delbrck Center for Molecular ... Research Center (ECRC) of MDC and Charit in ... of epithelial cells and their molecular repertoire. Dr. ... have identified a transcription factor (grainyhead-like 2, Grhl2), ...
Cached Biology News:Imperial and NTU's new medical school aspires to be global health care role model 2Imperial and NTU's new medical school aspires to be global health care role model 3Imperial and NTU's new medical school aspires to be global health care role model 4A wiki for the biofuels research community 2A wiki for the biofuels research community 3A wiki for the biofuels research community 4New insights into the development of epithelial cells 2
... D-2000, but comes with a shutter (controlled via ... D-2000 Deuterium Light Source produces a powerful, ... versions of the light source are available, providing ... and stable lamp provides peak-to-peak stability of less ...
... CYCLES. An assay kit designed to detect ... PARP is an enzyme implicated in DNA ... is cleaved by caspase-3. Cleavage of PARP ... kDa is a hallmark of apoptosis. Each ...
... These handy dispensers save space ... wipers, hats, and other garb. ... storage bins for small parts ... manufacturing. Clear acrylic. Open-slot design ...
... is expressed in g per ml and is ... pH 6.8, of a 1 mg per ml ... concentration to agglutinate a 2% suspension of human ... at 25 C. Preparation Note: Affinity ...
Biology Products: