Thermo Scientific Syn-PER Synaptic Protein Extraction Reagent is a proprietary cell lysis reagent for efficient isolation of synaptosomes containing functional synaptic proteins from neuronal tissue and primary cultured neurons.
Syn-PER Reagent is used to prepare synaptosomes containing biologically active pre-and post-synaptic proteins (i.e., intact membranes and protein complexes of synapses). When used with fresh neuronal tissue or primary cultured neurons, synaptosomes prepared with the Syn-PER Reagent can be used to study synaptic transmission. The synaptosomal proteins contained in these extracts can also be used for downstream applications such as Western blotting, immunoprecipitation, enzymatic activity assays and protein-protein interaction studies. Syn-PER Synaptic Protein Extraction Reagent minimizes the degradation of phosphoproteins and is ideal for studies requiring the preservation of phosphoprotein integrity.
- Efficient synapse extraction – obtain up to 10µg of synaptic protein per milligram of neuronal tissue or 4µg synaptic protein per 35mm dish of primary cultured neurons (106 cells)
- Gentle formulation – isolate viable synaptosomes; then lyse these synapses to extract native synaptic proteins and preserve phosphoprotein integrity
- Fast procedure – obtain synaptosomal suspension of intact synapses in less than one hour
- Simple protocol – requires no ultracentrifugation steps
- Isolate functional synaptosomes to study neurotransmitter release
- Extract pre- and post-synaptic proteins to identify changes in protein composition and function in synapses
- Preserve and study labile or transient protein phosphorylation events associated with synapses
Synaptosomes are isolated nerve terminals that are generated during the homogenization of nerve tissue. Synaptosome preparations can be used for extraction of pre- and post-synaptic proteins because they contain the complete pre-synaptic terminal, including mitochondria and synaptic vesicles, along with the post-synaptic membrane and the post-synaptic density. Synaptosomes are commonly used to study synaptic function because they contain functional ion channels, receptors, enzymes and intact synaptic vesicles, which can take up and release neurotransmitters. Synaptosome preparation typically involves several ultracentrifugation steps and density gradients to separate this cell fraction from other organelles.
Syn-PER Synaptic Protein Extraction Reagent efficiently enriches pre-and post-synaptic protein within synaptosomes with high yield. The nondenaturing cell lysis reagent is compatible with many downstream applications, including neurotransmitter release assays, enzyme assays (e.g., phosphatase, kinases), immunoassays, various chromatography procedures, and electrophoresis. In addition, Syn-PER Reagent preserves phosphoprotein integrity better than most commercially available extraction buffers, even in the absence of phosphatase inhibitors.
The Syn-PER Reagent does not contain protease or phosphatase inhibitors. However, inhibitors such as Thermo Scientific Halt Protease and/or Phosphatase Inhibitors can be added just before use to prevent proteolysis or to offer additional protection from the high phosphatase activity normally present in brain tissue.
|Greater enrichment of synaptic proteins is achieved in samples prepared using the Thermo Scientific Syn-PER Reagent than a homebrew reagent. Total protein (10µg) from mouse brain tissue homogenates (H), cytosol (C) fraction, and synaptosome suspension (Syn) were analyzed by Western blot. The total protein yield in the synaptosome suspension produced from fresh mouse brain (200mg) was three-fold greater using the Syn-PER Reagent compared to a typical homebrew reagent.
Review of cell lysis and protein extraction
N-PER Neuronal Protein Extraction Reagent
Other cell lysis and fractionation reagents
Protease and Phosphatase Inhibitor Cocktails and Tablets
Pierce Tissue Strainers