HEPES
Company: Sigma-Aldrich
Catalog#: H3375
Product evaluation:
FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction
Author:
Date:
2017-09-05
[Abstract]  We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the Drosophila neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as during the spontaneous release mode. The preparations are loaded with the FM1-43 dye, pre-fixed, treated and illuminated to photoconvert the dye, and then processed for conventional electron microscopy. This ...

Ubiquitin Proteasome Activity Measurement in Total Plant Extracts
Author:
Date:
2017-09-05
[Abstract]  The fine-tuned balance of protein level, conformation and location within the cell is vital for the dynamic changes required for a cell to respond to a given stimulus. This requires the regulated turnover of damaged or short-lived proteins through the ubiquitin proteasome system (UPS). Thus, the protease activity of the proteasome is adjusted to meet the current demands of protein degradation via the UPS within the cell. We describe the adaptation of an intramolecular quenched fluorescence ...

Preparation of Crude Synaptosomal Fractions from Mouse Brains and Spinal Cords
Author:
Date:
2017-08-05
[Abstract]  The current protocol describes the preparation of crude synaptosomal fractions from mouse brain or spinal cord samples. In detail, a sequential protocol yielding crude synaptosomal and light membrane fractions is provided. This fast and easy method might be sufficient to assess the amount of synaptic proteins in down-steam applications like Western-blot or ELISA in e.g., mouse models of Alzheimer’s disease or other neurodegenerative conditions.

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