| Autor: |
Marie Kelly-Worden, Sara Ghassemifar |
| Rok vydání: |
2008 |
| Předmět: |
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| Zdroj: |
Advances in Planar Lipid Bilayers and Liposomes ISBN: 9780123743411 |
| Popis: |
Artificial vesicles have been used for decades to examine reconstituted purified membrane proteins and reconstitution of pieces of cellular (mCs) or vesicular membranes (mVs). Reconstitution of vesicles into liposomes allows vesicular proteins to maintain a semi‐natural environment. These mVs can then be incorporated into planar lipid bilayers either by direct brushing or by incorporating through the use of an osmotic gradient. Incorporation can be monitored by the nystatin/ergosterol technique or by observing capacitance change. Liposomes and mVs can also be used for examining autoimmune antibodies–vesicular membranes interaction using poly‐ d ‐lysine plated coverslips. Modified synaptic vesicles (mSVs) can be prepared and plated onto poly‐ d ‐lysine coated coverslips. The mSVs can be processed using primary antibodies against specific synaptic vesicular proteins (such as VAMP and Rabphilin‐3A) and the appropriate secondary Alexa fluor antibody. Simultaneously the mSVs can be examined for the ability of autoimmune antibodies to bind with these vesicles, and any cross‐reactivity can be observed by fluorescent microscopy. Here we describe fluorescent techniques for examining liposomes, synaptic vesicles, and mSVs as well as bilayer techniques for examining mSVs. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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