scholarly journals Determination of Membrane Protein Transporter Oligomerization in Native Tissue Using Spatial Fluorescence Intensity Fluctuation Analysis

PLoS ONE ◽  
2012 ◽  
Vol 7 (4) ◽  
pp. e36215 ◽  
Author(s):  
Mikhail Sergeev ◽  
Antoine G. Godin ◽  
Liyo Kao ◽  
Natalia Abuladze ◽  
Paul W. Wiseman ◽  
...  
2010 ◽  
Vol 98 (3) ◽  
pp. 575a-576a
Author(s):  
Mikhail Sergeev ◽  
Antoine G. Godin ◽  
Liyo Kao ◽  
Natalia Abuladze ◽  
Ira Kurtz ◽  
...  

2009 ◽  
Vol 96 (3) ◽  
pp. 34a
Author(s):  
Mikhail Sergeev ◽  
Antoine G. Godin ◽  
Jody L. Swift ◽  
Yves De Koninck ◽  
Paul W. Wiseman

2019 ◽  
Author(s):  
Michael R. Stoneman ◽  
Gabriel Biener ◽  
Richard J. Ward ◽  
John D. Pediani ◽  
Dammar Badu ◽  
...  

Abstract Current methods for determining membrane protein association in cells are either very time consuming, require complicated procedures, or lack the sensitivity needed to assess the effect of concentration or ligand binding on the observed oligomerization. To overcome these limitations, we provide a detailed protocol for quantifying the relative abundance and stability of oligomers of differing sizes using two-dimensional fluorescence intensity fluctuation (2D-FIF) spectrometry. The approach can be implemented using a standard laser scanning fluorescence microscope along with custom written software for image analysis. This method may be applied to evaluate the extent of oligomerization as a function of receptor concentration and is particularly suited to assess the effects of agonists and antagonists on the oligomeric size of membrane receptors of interest.


2010 ◽  
Vol 24 (S1) ◽  
Author(s):  
Mikhail Sergeev ◽  
Antoine G Godin ◽  
Liyo Kao ◽  
Natalia Abuladze ◽  
Paul W Wiseman ◽  
...  

Molecules ◽  
2021 ◽  
Vol 26 (5) ◽  
pp. 1246
Author(s):  
Tengfei Wang ◽  
Hui Luo ◽  
Xu Jing ◽  
Jiali Yang ◽  
Meijun Huo ◽  
...  

Water-soluble fluorescent carbon dots (CDs) were synthesized by a hydrothermal method using citric acid as the carbon source and ethylenediamine as the nitrogen source. The repeated and scale-up synthetic experiments were carried out to explore the feasibility of macroscopic preparation of CDs. The CDs/Fe3+ composite was prepared by the interaction of the CDs solution and Fe3+ solution. The optical properties, pH dependence and stability behavior of CDs or the CDs/Fe3+ composite were studied by ultraviolet spectroscopy and fluorescence spectroscopy. Following the principles of fluorescence quenching after the addition of Fe3+ and then the fluorescence recovery after the addition of asorbic acid, the fluorescence intensity of the carbon dots was measured at λex = 360 nm, λem = 460 nm. The content of ascorbic acid was calculated by quantitative analysis of the changing fluorescence intensity. The CDs/Fe3+ composite was applied to the determination of different active molecules, and it was found that the composite had specific recognition of ascorbic acid and showed an excellent linear relationship in 5.0–350.0 μmol·L−1. Moreover, the detection limit was 3.11 μmol·L−1. Satisfactory results were achieved when the method was applied to the ascorbic acid determination in jujube fruit. The fluorescent carbon dots composites prepared in this study may have broad application prospects in a rapid, sensitive and trace determination of ascorbic acid content during food processing.


1990 ◽  
Vol 10 (2) ◽  
pp. 131-139
Author(s):  
Oyewole Adeyemo ◽  
E. O. Okegbile ◽  
O. O. Olorunsogo

For the development of immunological contraception, attention is being concentrated on the possibility of using a sperm membrane antigen. Boar sperm membrane was extracted with triton-X 100 and fractionated by Sephadex G-150 column chromatography. The glycosylated and nonglycosylated portions of protein peaks from the gel filtration were obtained by fractionating on concanavalin A-Sepharose and eluting the bound protein with 0.3 M methyl mannoside. A glycosylated fraction was found to induce sperm agglutinating antibodies in rabbit. The partially purified protein has a molecular weight of 30 kilodaltons, as determined by sodium dodecyl polyaccyrlamide gel electrophoresis. Further work is planned on the histochemical determination of the origin of this protein and species cross-activity of the antibody.


2005 ◽  
Vol 128 (2) ◽  
pp. 203-209 ◽  
Author(s):  
L. Zhu ◽  
Y. Zheng ◽  
C. H. von Kerczek ◽  
L. D. T. Topoleski ◽  
R. W. Flower

Indocyanine green (ICG) dye angiography has been used by ophthalmologists for routine examination of the choroidal vasculature in human eyes for more than 20years. In this study, a new approach is developed to extract information from ICG dye angiograms about blood velocity distribution in the choriocapillaris and its feeding blood vessels. ICG dye fluorescence intensity rise and decay curves are constructed for each pixel location in each image of the choriocapillaris in an ICG angiogram. It is shown that at each instant of time the magnitude of the local instantaneous dye velocity in the choriocapillaris is proportional to both the slope of the ICG dye fluorescence intensity curve and the dye concentration. This approach leads to determination of the absolute value of blood velocity in the choriocapillaris, assuming an appropriate scaling, or conversion factor can be determined. It also enables comparison of velocities in different regions of the choriocapillaris, since the conversion factor is independent of the vessel location. The computer algorithm developed in this study can be used in clinical applications for diagnostic purposes and for assessment of the efficacy of laser therapy in human eyes.


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