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Fibulin-1 Purification From Human Plasma Using Affinity Chromatography on Factor H-Sepharose

Fibulin-1 Purification From Human Plasma Using Affinity Chromatography on Factor H-Sepharose

Richard G DiScipio, Robert C Liddington, Ingrid U Schraufstatter

Protein Expr Purif. 2016 May;121:118-24.

PMID: 26826315

Abstract:

A method is reported to purify Fibulin-1 from human plasma resulting in a 36% recovery. The steps involve removal of the cryoglobulin and the vitamin K dependent proteins followed by polyethylene glycol and ammonium sulfate precipitations, DEAE-Sephadex column chromatography and finally Factor H-Sepharose affinity purification. The procedure is designed to be integrated into an overall scheme for the isolation of over 30 plasma proteins from a single batch of human plasma. Results from mass spectroscopy, SDS-PAGE, and Western blotting indicate that human plasma Fibulin-1 is a single chain of the largest isotype. Functional binding assays demonstrated calcium ion dependent interaction of Fibulin-1 for fibrinogen, fibronectin, and Factor H. The procedure described is the first to our knowledge that enables a large scale purification of Fibulin-1 from human plasma.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP80295654	Complement factor H from human plasma		80295-65-4	Price
AP9001325	Fibrinogen from human plasma		9001-32-5	Price

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