Low background metal ion slides for poly-His tagged protein molecules

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Advanced Surface Coating Technologies for Microarrays & Microfluidics

	Zero background copper slides for the selective immobilization of poly-His tagged protein molecules with controlled orientation!

	Immobilization of proteins without purification
		Comparison of immunostaining of sulfotransferases with and without purification (A) an array of purified 6x His-Sulfotransferase IV (His-ST IV) on the Zero Background copper slide; (B) background test of purified His-ST IV on the Zero Background slide without Cu2+ activation; (C) an array of crude lysates containing His-ST IV on the Zero Background copper slide; (D) background test for His-ST IV in crude lysates on the Zero Background slide without Cu2+ activation. Protein detection was achieved by incubation with primary antibody against ST IV, followed by Cy3-labeled secondary antibody.
		Left: Immobilization of 6xHis-tagged green fluorescent protein (His-GFP, not purified) on the Zero Background copper slide. Right: Background test for GFP on the same surface.
	Superior advantage of the oriented protein array: a comparison to randomly oriented protein molecules on the surface



	Activities 6xHis-tagged sulfotransferase for (A&B) randomly oriented proteins on the surface; (C) oriented proteins on the zero-background Cu2+ surface; and (D) native state in the solution phase.	Activities 6xHis-alkaline phosphatase for (A) randomly oriented proteins on the surface; (B) oriented proteins on the zero-background Cu2+ surface; and (C) native state in the solution phase.
	Only oriented protein molecules faithfully reflect activities of solution phase proteins!
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