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Fluorimetric Studies and Noncovalent Labeling of Protein With the Near-Infrared Dye HITCI for Analysis by CE-LIF

Fluorimetric Studies and Noncovalent Labeling of Protein With the Near-Infrared Dye HITCI for Analysis by CE-LIF

Weiying Yan, Christa L Colyer

J Sep Sci. 2005 Aug;28(12):1409-15.

PMID: 16138693

Abstract:

1,1',3,3,3',3'-Hexamethylindotricarbocyanine iodide (HITCI) is a commercially available, positively charged, indocarbocyanine dye used typically as a laser dye in the near infrared (NIR). The absorbance and fluorescence properties of HITCI in a variety of solvent systems were determined. Results indicate that the fluorescence of HITCI is not significantly affected by the pH. Titration of HITCI with human serum albumin (HSA) and trypsinogen was carried out to investigate the interactions between this dye and proteins. These studies revealed that the absorbance and fluorescence properties of the dye change upon binding to protein in a wide range of solution pH's. The potential use of HITCI as a noncovalent protein labeling probe, therefore, was explored. Determination and separation of HITCI and HITCI-protein complexes was performed by capillary electrophoresis with diode-laser induced fluorescence detection (CE-LIF). Both pre-column and on-column noncovalent labeling methods are demonstrated.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP19764966	1,1′,3,3,3′,3′-Hexamethylindotricarbocyanine iodide		19764-96-6	Price

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