Navigation Links
Purification of Phosphoinositide 3-Kinase With UNO Columns


Polyphosphoinositides and their metabolites are important intracellular signals involved in the responses to a number of hormones and growth factors.The discovery of a phosphatidylinositol kinase that phosphorylates PI at the 3 position of the inositol ring uncovered a new pathway of PI metabolism and potential intracellular signals.

Phosphoinositide 3-kinase has been purified to homogeneity from rat liver and bovine thymus. The enzyme from rat liver is a heterodimer of 110 and 85 kDa proteins.Two forms of the 110 kDa protein exist, and have similar but not identical amino acid sequences. Recent studies have suggested that the 110 kDa protein is the catalytic protein and that the 85 kDa protein plays a regulatory role.

Phosphoinositide 3-kinase has proven difficult to purify in an active form when overexpressed in various expression systems. Consequently, for continued biochemical studies of this phosphoinositide pathway, a routine purification protocol for phosphoinositide 3-kinase from rat liver has been established.The protocol involves multiple steps including size exclusion, hydroxyapatite, and ion-exchange chromatography.

The chromatograms (Figures 1 and 2) illustrate the use of an UNO Q1 column to partially purify a cytosolic phosphoinositide 3-kinase from a rat liver homogenate at a flow rate 5-fold higher than previously used with a conventional monodispersed beaded media.

Both the overall resolution and protein activity profile are maintained at the higher flow.


Acknowledgement
Data courtesy of Dr.A. Couvillon, Beth Israel Deaconess Medical Center, Boston, MA.


FPLC is a trademark of Pharmacia


back to top
'"/>

Source:


Page: All 1 2

Related biology technology :

1. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
2. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
3. Yeast Protein Production System Features High Yields and One-Step Purification
4. Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
5. Easy Cosmid DNA Purification with StrataPrep Kits
6. MinElute PCR Purification Kit
7. QIAquick PCR Purification Kit
8. Microarray Target Purification Kit and the Microarray Target Synthesis Kit (T7)
9. Double-Affinity Purification of GST-His Tagged Proteins Using Vivapure 8-to-96 Well Cobalt-Chelate Kit
10. Ultra-Fast Purification of Small Nucleic Acids
11. RNAClean - RNA & cDNA in vitro Reaction Purification
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:12/6/2016)... ... December 06, 2016 , ... ... announced today that the company has engaged in a collaborative research partnership with ... with the CSU Office of the Vice President for Research. This agreement is ...
(Date:12/6/2016)... Dec. 6, 2016  SRI International has been ... from the National Institutes of Health,s National Institute ... of AIDS (NIAID-DAIDS) to support the manufacturing and ... (PreP) agents. Under the seven-year contract, SRI will ... services for candidate HIV-prevention products that emerge from ...
(Date:12/5/2016)... (PRWEB) , ... December 05, 2016 , ... ... nanocellulose, including both cellulose nanocrystals and cellulose nanofibrils. The composition claims are ... There are also claims directed to combination with polymers, carbon fibers, graphene, and ...
(Date:12/5/2016)... 5, 2016  Renova™ Therapeutics, a biotechnology company ... and other chronic diseases, announced that Catherine ... Chief Financial Officer (CFO), effective today. ... of experience in financial management for a variety ... companies. Most recently, Ms. Bovenizer was the Vice ...
Breaking Biology Technology:
(Date:11/30/2016)... FRANCISCO and WARSAW, Poland , Nov. 30, 2016 /PRNewswire/ ... It is one of the most crucial aspects of recovery so we need to ... of serious health risks, including heart problems, high blood pressure, stroke, diabetes, and even ... and friends sleep and find a Christmas present that could help them to manage ... ...
(Date:11/29/2016)... VILNIUS, Lithuania , Nov. 29, 2016 ... high-precision biometric identification and object recognition technologies, ... kit (SDK) for fingerprint recognition solutions that ... extract a fingerprint template using less than ... used in compact devices that have limited ...
(Date:11/24/2016)... IRVINE, Calif. , Nov. 23, 2016 ... help endurance athletes and their trainers non-invasively measure ... Variability Index, Pulse Rate, and Respiration Rate in approximately ... Ember enables users easy and immediate access to key ... as part of a training regimen. ...
Breaking Biology News(10 mins):