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Measurements of interactions of memb...

Hagen, Guy M.

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Measurements of interactions of membrane proteins.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Measurements of interactions of membrane proteins./
作者:	Hagen, Guy M.
面頁冊數:	121 p.
附註:	Source: Dissertation Abstracts International, Volume: 66-08, Section: B, page: 4196.
Contained By:	Dissertation Abstracts International66-08B.
標題:	Chemistry, Analytical. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3185507
ISBN:	9780542268441

Measurements of interactions of membrane proteins.
Hagen, Guy M.

Measurements of interactions of membrane proteins. - 121 p.

Source: Dissertation Abstracts International, Volume: 66-08, Section: B, page: 4196.

Thesis (Ph.D.)--Colorado State University, 2005.

We have improved upon conventional fluorescence photobleaching recovery (FPR) measurements by introducing two new techniques: total internal reflection interference fringe (TIRIF)-FPR, and high probe intensity (HPI)-FPR. Both of these techniques are designed to enhance diffusion measurements of visible fluorescent protein (VFP)-membrane receptor fusion proteins. TIRIF-FPR restricts photoexcitation to the cell membrane, eliminating contributions to the recovery signal from fluorescent cytoplasmic species. HPI-FPR accomplishes measurements of sparsely-expressed VFP-fusion proteins where conventional measurements fail by increasing laser power in spot-FPR methods. Data with increased fluorescent probe photobleaching in the attenuated measurement beam are thus recorded, and for the first time, correctly evaluated.

ISBN: 9780542268441Subjects--Topical Terms:

586156
Chemistry, Analytical.

Measurements of interactions of membrane proteins.
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500 $a Source: Dissertation Abstracts International, Volume: 66-08, Section: B, page: 4196.
500 $a Adviser: George Barisas.
502 $a Thesis (Ph.D.)--Colorado State University, 2005.
520 $a We have improved upon conventional fluorescence photobleaching recovery (FPR) measurements by introducing two new techniques: total internal reflection interference fringe (TIRIF)-FPR, and high probe intensity (HPI)-FPR. Both of these techniques are designed to enhance diffusion measurements of visible fluorescent protein (VFP)-membrane receptor fusion proteins. TIRIF-FPR restricts photoexcitation to the cell membrane, eliminating contributions to the recovery signal from fluorescent cytoplasmic species. HPI-FPR accomplishes measurements of sparsely-expressed VFP-fusion proteins where conventional measurements fail by increasing laser power in spot-FPR methods. Data with increased fluorescent probe photobleaching in the attenuated measurement beam are thus recorded, and for the first time, correctly evaluated.
520 $a We have compared both new and existing FPR techniques on a variety of biological systems including measurements of the mast cell function-associated antigen (MAFA). This regulatory protein is involved in IgE-receptor mediated allergic responses and was also studied using time-resolved phosphorescence anisotropy (TPA) methods. TPA results, which are uniquely sensitive to in-membrane molecular size, indicate the IgE receptor and the MAFA regulatory protein are linked in the membrane under both resting and activating conditions.
520 $a As part of an on-going effort to improve TPA measurements, new strategies for photomultiplier tube (PMT) gating have also been devised. Saturation of the photomultiplier used in TPA measurements during the intense visible laser flash is a serious problem which can cause artifactual signals and damage to the detector. This was previously addressed by rapidly gating the detector off, but artifactual signals arising at the end of the off-gated period persisted. We have thus developed a fast, fully adjustable photomultiplier gate with high extinction that is capable of eliminating the light-induced post gate artifacts encountered with earlier gating techniques.
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