Yan Weiying, Colyer Christa L
Wake Forest University, Department of Chemistry, Winston-Salem, NC 27109, USA.
J Sep Sci. 2005 Aug;28(12):1409-15. doi: 10.1002/jssc.200500024.
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.
1,1',3,3,3',3'-六甲基吲哚三碳菁碘化物（HITCI）是一种市售的带正电荷的吲哚碳菁染料，通常用作近红外（NIR）激光染料。测定了HITCI在多种溶剂体系中的吸光度和荧光特性。结果表明，HITCI的荧光不受pH值的显著影响。用人类血清白蛋白（HSA）和胰蛋白酶原对HITCI进行滴定，以研究该染料与蛋白质之间的相互作用。这些研究表明，在广泛的溶液pH值范围内，该染料与蛋白质结合后，其吸光度和荧光特性会发生变化。因此，探索了HITCI作为非共价蛋白质标记探针的潜在用途。采用二极管激光诱导荧光检测毛细管电泳（CE-LIF）对HITCI和HITCI-蛋白质复合物进行测定和分离。展示了柱前和柱上非共价标记方法。
