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.

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

FPLC is a trademark of Pharmacia

back to top


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:

(Date:11/24/2015)... DIEGO , Nov. 24, 2015 Halozyme Therapeutics, Inc. ... Healthcare Conference in New York on Wednesday, ... Helen Torley , president and CEO, will provide a corporate ... New York at 1:00 p.m. ET/10:00 a.m. PT ... and investor relations, will provide a corporate overview. --> ...
(Date:11/24/2015)... /PRNewswire/ - Aeterna Zentaris Inc. (NASDAQ:  AEZS) (TSX: AEZ) (the ... the Toronto Stock Exchange, confirms that as of the ... developments that would cause the recent movements in the ... --> About Aeterna Zentaris Inc. ... Aeterna Zentaris is a specialty biopharmaceutical company engaged in ...
(Date:11/24/2015)... ... ... The Academy of Model Aeronautics (AMA), led by its Executive Council, has officially ... to represent the First–Person View (FPV) racing community. , FPV racing has exploded in ... racing and several new model aviation pilots have joined the community because of their ...
(Date:11/24/2015)... , Nov. 24, 2015 /PRNewswire/ - Aeterna ... announced today that the remaining 11,000 post-share consolidation ... Purchase Warrants (the "Series B Warrants") subject to ... exercised on November 23, 2015, which will result ... After giving effect to the issuance of such ...
Breaking Biology Technology:
(Date:10/26/2015)... , October 26, 2015 /PRNewswire/ ... --> adds Biometrics ... to 2021 as well as Emerging ... research reports to its collection of ... . --> ...
(Date:10/23/2015)... 23, 2015 Research and Markets ( ... Voice Recognition Biometrics Market 2015-2019" report to their ... The global voice recognition biometrics market to grow ... --> --> The report, ... based on an in-depth market analysis with inputs from ...
(Date:10/22/2015)... JOSE, Calif., Oct. 22, 2015  Synaptics (NASDAQ: SYNA ), ... for its first quarter ended September 30, 2015. ... quarter of fiscal 2016 grew 66 percent over the comparable quarter ... of fiscal 2016 was $23.8 million, or $0.62 per diluted share. ... income for the first quarter of fiscal 2016 grew 39 percent ...
Breaking Biology News(10 mins):