Navigation Links
DELFIA Protein Kinase assays.

Protein kinases use ATP to phosphorylate certain amino acid residues (tyrosine, serine or threonine) on their substrates. In receptormediated signalling the phosphorylation forms an important secondary messenger system. Phosphorylation can be measured following radioactive 32P incorporation, which, however, involves the problems of radioactive work, safety and disposal. Eu-labelled anti-phosphotyrosine, -threonine and threonine-proline are available for the sensitive detection of phosphorylation levels on various substrates using time-resolved fluorometry and DELFIA as the detection technology.

PROTOCOL

Convenient collection of the substrates (peptides, polypeptides or proteins) on coated microtitration plates can be accomplished using streptavidin plates for biotinylated substrates or glutathione or anti-glutathione-S-transferase plates for substrates expressed with a GST tag.

1. In the assay, first incubate the substrate with enzyme (receptor), ATP and effectors.

2. After phosphorylation collect the phosphorylated products on coated plates. Incubate 1-2 hours and wash the plate two or three times with wash solution.

3. Add Eu-(or biotin-) labelled antibodies in DELFIA Assay Buffer (200 μl, 250 ng/ml) and incubate for 1-2 h. If performed in the indirect way, the secondary incubation with Eu-streptavidin takes 20-30 minutes.

4.Wash the plate six times with wash solution using the DELFIA Platewash (1296-024 or 026).

5. Add 200 μl of DELFIA Enhancement Solution, shake for 10 min with Plateshaker (1296-001/002 or 003/004) to guarantee a stable signal level and measure the plate with the Wallac time-resolved fluorometer.

REFERENCES

Assays are currently routinely used in a number of drug discovery laboratories
'"/>

Source:


Page: All 1 2

Related biology technology :

1. A DELFIA Assay for Inhibition of PARP
2. A New C-Terminal GST Vector for Protein Production in S. pombe
3. New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
4. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
5. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
6. New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
7. Optimized Imaging of Protein Gels Stained with Coomassie Brilliant Blue Dye
8. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
9. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
10. Yeast Protein Production System Features High Yields and One-Step Purification
11. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: DELFIA Protein Kinase assays

(Date:5/21/2013)... -- Synthetic Biologics, Inc. (NYSE MKT: SYN), a developer of biologics ... announced today that Jeffrey Riley , CEO, ... MicroCap Conference on Thursday, May 30, 2013 in ... Mr. Riley is scheduled to present at 10:00 a.m. (Eastern ... can be accessed by logging onto the web at ...
(Date:5/21/2013)... 21, 2013 Inflamax scientists worked ... ITN to design the study, which will provide ... clinical models used in the study of allergy ... Allergen Challenge (NAC) models. Although both the models ... to date no direct comparison between the allergic ...
(Date:5/21/2013)... MN (PRWEB) May 21, 2013 The ... show display solution for sales and marketing teams looking for ... describes the kit as “everything you’ll need to put ... Fabric Display Kit comes with a number of displays and ... trade booth 20 feet by 20 feet in size. These ...
(Date:5/20/2013)...  MiMedx Group, Inc. (NASDAQ: MDXG ), ... regenerative biomaterials and bioimplants processed from human amniotic membrane, ... of credit with Bank of America.  The revolving line ... accounts receivable and inventory, allows MiMedx to borrow up ... will be used for general corporate purposes. ...
Breaking Biology Technology:Synthetic Biologics to Present at the 2nd Annual Marcum LLP MicroCap Conference 2Inflamax Research selected by the Immune Tolerance Network and the National Institutes of Health to conduct a landmark clinical study on the underlying mechanisms of allergic inflammation. 2GLM Displays Announces All-in-One Formulate Fusion Fabric Display Kit 2
... LAFAYETTE, Ind. - Researchers at Purdue University are ... fit inside laptops and personal computers, a cooling ... size of computers. , Unlike conventional cooling systems, ... finned devices called heat sinks attached to computer ...
... JumpStart Inc., the Northeast Ohio,venture development organization ... and ideas, has announced an investment commitment ... JumpStart,investment is part of a recently closed ... Management of Cleveland, OH., (Logo: ...
... to Adipose-Derived Stem Cells; business ... likely affected., SUNNYVALE, Calif., June 19 ... concerning the foundational,composition-of-matter patent covering stem cells isolated ... of Artecel,s licensor, the,University of Pittsburgh (,Pittsburgh,) and ...
Cached Biology Technology:Tiny refrigerator taking shape to cool future computers 2Tiny refrigerator taking shape to cool future computers 3JumpStart Invests in Tursiop Technologies 2JumpStart Invests in Tursiop Technologies 3Court Determines that University of Pittsburgh and Artecel Have Sole Rights to Adipose-Derived Stem Cell Patent 2Court Determines that University of Pittsburgh and Artecel Have Sole Rights to Adipose-Derived Stem Cell Patent 3Court Determines that University of Pittsburgh and Artecel Have Sole Rights to Adipose-Derived Stem Cell Patent 4
(Date:5/21/2013)... battle? Try staying fit in space, where living quarters ... , That,s the challenge a group of Michigan State ... million grant from NASA. Their goal is to keep ... "You could give astronauts the best exercise regimen there ... to it, it isn,t going to help," said project ...
(Date:5/21/2013)... ANDOVER, Mass. , May 21, 2013 ... that CX50 xMATRIX, the world,s first portable ultrasound with ... Intracardiac Echo (ICE) capability. The CX50 xMATRIX with available ... Paris at the EuroPCR, official ... (EAPCI), May 21-24. Since its launch in ...
(Date:5/20/2013)... the lungs of the planet, inhales carbon dioxide as it ... grow parts that eventually fall to the ground to decompose ... , Until recently people believed much of the rain forest,s ... in the ocean. University of Washington research showed a decade ... though left open the question of how that was possible, ...
Breaking Biology News(10 mins):Fueling fitness on the final frontier 2Philips CX50 xMATRIX now offers world-class interventional and diagnostic features on single portable system 2Amazon River exhales virtually all carbon taken up by rain forest 2Amazon River exhales virtually all carbon taken up by rain forest 3
... a potentially patentable method to arrest toxic leakages of ... gene therapy against cancerous tumors. The problem has been ... from tumors, proving toxic to other body tissues. , ... changes from a liquid at 39 degrees Fahrenheit to ...
... The recent discovery of a 90-million-year-old dinosaur in ... theropods that includes Velociraptor and is closely related ... Rather than originating during the Cretaceous, dromaeosaurs can ... as far back as 180 million years ago. ...
... exact chemical blend of a pheromone emitted by older male ... mating and how other surrounding elephants behave, a new study ... Science University and the University of Auckland in New Zealand ... images of the pheromone, frontalin, depends on whether the male ...
Cached Biology News:Polymer gel can block toxic leakage problem in gene therapy 2Polymer gel can block toxic leakage problem in gene therapy 3Polymer gel can block toxic leakage problem in gene therapy 4Newly discovered birdlike dinosaur is oldest raptor ever found in South America 2Newly discovered birdlike dinosaur is oldest raptor ever found in South America 3Newly discovered birdlike dinosaur is oldest raptor ever found in South America 4Male elephants woo females with precise chemistry 2Male elephants woo females with precise chemistry 3
...
Removes polysaccharide and phenolic inhibitors of PCR; yields DNA greater than 50 kb in length...
...
Caspase Activity Detector, SR-VAD-FMK Kit , 100 tests Consult technical datasheet for details....
Biology Products: