Indentification and detection of cis-platin binding proteins by laser induced breakdown spectroscopy için kapak resmi
Indentification and detection of cis-platin binding proteins by laser induced breakdown spectroscopy
Kaya, İbrahim, author.
Yazar Ek Girişi:
Fiziksel Tanımlama:
x, 56 leaves.: + 1 computer laser optical disc.
In this study, an all-optically designed laser plasma spectroscopic technique for rapid identification and detection of cisplatin-binding proteins on electrophoretic gel spots prior to molecular mass spectrometric analysis is demonstrated. For this purpose, human serum albumin, human apo transferrin and horse heart myoglobin standard proteins and protein extracts from HeLa cancer cells were subjected to; incubation with cis-platin solution for several hours. Then, non-reducing polyacrylamide gel electrophoretic separation was applied. Followed by the visualization of proteins in the gel by Coomassie Brilliant Blue staining technique protein spots on the gel were dried between two cellophane sheets and subjected to laser ablation by highly energetic laser pulses. In addition, prior to nr-SDS-PAGE separation cis-platin binding to standard proteins were monitored by ESI-MS with several measurements made in 24 hours of incubation time. Using a Nd:YAG laser at its second harmonic wavelength, 532nm, 10 Hz frequency and 10 ns pulse duration, a micro-plasma was created on dried gel spots. Resulting plasma emission light was collected with collection lenses and transferred to a spectrograph via fiber optic cable. An intensified charge coupled device (ICCD) detector enabled multielemental analysis of platinum binding protein samples. Platinum binding proteins were recognized from the prominent neutral emission line, Pt (I) at 273.3 nm, in a plasma formed by the focused laser pulses on the gel, just in the center or in the vicinity of the electrophoretic spot. Spectral emission intensity of Pt lines from LIBS data has been optimized with respect to laser energy and detector timing parameters. Optimization of LIBS experimental parameters have been studied on polyacrylamide gels soaked in cis-Pt solution for Pt signal. It has been shown that, LIBS is a suitable method for identifying Pt in proteins, in gel medium, with nanogram levels of detection capability. The technique was applied to HeLa (human cervical cancer cells) cells extract for the detection of Pt-binded HSA after standard addition of known amounts of protein.
Yazar Ek Girişi:
Tek Biçim Eser Adı:
Thesis (Master)--İzmir Institute of Technology:Chemistry.

İzmir Institute of Technology:Chemistry--Thesis (Master.
Elektronik Erişim:
Access to Electronic Versiyon.


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Durumu/İade Tarihi
Tez T001417 QD96.P62 K23 2015

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