A cell-free assay to determine the stoichiometry of plasma membrane proteins
Plasma membrane receptors, transporters, and ion channel molecules are often found as oligomeric structures that participate in signaling cascades essential for cell survival. Different states of protein oligomerization may play a role in functional control and allosteric regulation. Stochastic GFP-...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2013-04-01
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| Series: | BioTechniques |
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| Online Access: | https://www.future-science.com/doi/10.2144/000113977 |
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| _version_ | 1850152158879547392 |
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| author | Cesar Trigo Juan P Vivar Carlos B Gonzalez Sebastian Brauchi |
| author_facet | Cesar Trigo Juan P Vivar Carlos B Gonzalez Sebastian Brauchi |
| author_sort | Cesar Trigo |
| collection | DOAJ |
| description | Plasma membrane receptors, transporters, and ion channel molecules are often found as oligomeric structures that participate in signaling cascades essential for cell survival. Different states of protein oligomerization may play a role in functional control and allosteric regulation. Stochastic GFP-photobleaching (SGP) has emerged as an affordable and simple method to determine the stoichiometry of proteins at the plasma membrane. This non-invasive optical approach can be useful for total internal reflection of fluorescence microscopy (TIRFM), where signal-to-noise ratio is very high at the plasma membrane. Here, we report an alternative methodology implemented on a standard laser scanning confocal microscope (LSCM). The simplicity of our method will allow for its implementation in any epifluorescence microscope of choice. |
| format | Article |
| id | doaj-art-aab3009d94214644bfdba2895e6d93da |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2013-04-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-aab3009d94214644bfdba2895e6d93da2025-08-20T02:26:03ZengTaylor & Francis GroupBioTechniques0736-62051940-98182013-04-0154419119610.2144/000113977A cell-free assay to determine the stoichiometry of plasma membrane proteinsCesar Trigo0Juan P Vivar1Carlos B Gonzalez2Sebastian Brauchi31Instituto de Fisiología, Facultad de Medicina, Universidad Austral de Chile, Campus Isla Teja, Valdivia, Chile1Instituto de Fisiología, Facultad de Medicina, Universidad Austral de Chile, Campus Isla Teja, Valdivia, Chile1Instituto de Fisiología, Facultad de Medicina, Universidad Austral de Chile, Campus Isla Teja, Valdivia, Chile1Instituto de Fisiología, Facultad de Medicina, Universidad Austral de Chile, Campus Isla Teja, Valdivia, ChilePlasma membrane receptors, transporters, and ion channel molecules are often found as oligomeric structures that participate in signaling cascades essential for cell survival. Different states of protein oligomerization may play a role in functional control and allosteric regulation. Stochastic GFP-photobleaching (SGP) has emerged as an affordable and simple method to determine the stoichiometry of proteins at the plasma membrane. This non-invasive optical approach can be useful for total internal reflection of fluorescence microscopy (TIRFM), where signal-to-noise ratio is very high at the plasma membrane. Here, we report an alternative methodology implemented on a standard laser scanning confocal microscope (LSCM). The simplicity of our method will allow for its implementation in any epifluorescence microscope of choice.https://www.future-science.com/doi/10.2144/000113977LSCMTIRFMStochastic-GFP photobleachingmembrane proteinsmembrane sheetHv1 |
| spellingShingle | Cesar Trigo Juan P Vivar Carlos B Gonzalez Sebastian Brauchi A cell-free assay to determine the stoichiometry of plasma membrane proteins BioTechniques LSCM TIRFM Stochastic-GFP photobleaching membrane proteins membrane sheet Hv1 |
| title | A cell-free assay to determine the stoichiometry of plasma membrane proteins |
| title_full | A cell-free assay to determine the stoichiometry of plasma membrane proteins |
| title_fullStr | A cell-free assay to determine the stoichiometry of plasma membrane proteins |
| title_full_unstemmed | A cell-free assay to determine the stoichiometry of plasma membrane proteins |
| title_short | A cell-free assay to determine the stoichiometry of plasma membrane proteins |
| title_sort | cell free assay to determine the stoichiometry of plasma membrane proteins |
| topic | LSCM TIRFM Stochastic-GFP photobleaching membrane proteins membrane sheet Hv1 |
| url | https://www.future-science.com/doi/10.2144/000113977 |
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