准备和存储
推荐的实验流程
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml .
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Synaptotagmin (p65) is an abundant synaptic vesicle protein that contains a single transmembrane region and two copies of an internal repeat that is homologous to the regulatory region of Protein Kinase C. It appears that synaptotagmin has a regulatory role in the synaptic vesicle pathway, particularly in vesicle docking and/or fusion with the plasmalemma. A model has been proposed to explain docking, activation, and fusion of synaptic vesicles with donor membranes. This model suggests that VAMP/synaptobrevin and synaptotagmin (vSNARE) on the synaptic vesicle, and SNAP-25 and syntaxin (tSNAREs) on the plasma membrane, interact to form a 7S complex. Two additional soluble proteins, αSNAP and NSF, are later added to the 7S complex, accompanied by the loss of synaptotagmin. The resulting 20S complex contains syntaxin, SNAP-25, VAMP, αSNAP, and NSF. Genetic studies in several species demonstrate that mutation or deletion of synaptotagmin results in a large decrease in Ca2+ triggered transmitter release. Mammalian synapses that lack synaptotagmin show a selective decrease in a fast component of release, suggesting that synaptotagmin is the Ca2+ sensor triggering exocytosis.
研发参考 (5)
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Duncan RR, Don-Wauchope AC, Tapechum S, Shipston MJ, Chow RH, Estibeiro P. High-efficiency Semliki Forest virus-mediated transduction in bovine adrenal chromaffin cells. Biochem J. 1999; 342(Pt 3):497-501. (Clone-specific: Western blot). 查看参考
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Liu Y, Fallon L, Lashuel HA, Liu Z, Lansbury PT Jr. The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinson's disease susceptibility. Cell. 2002; 111(2):209-218. (Clone-specific: Western blot). 查看参考
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Perin MS, Johnston PA, Ozcelik T, Jahn R, Francke U, Sudhof TC. Structural and functional conservation of synaptotagmin (p65) in Drosophila and humans. J Biol Chem. 1991; 266(1):615-622. (Biology). 查看参考
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Ramalho-Santos J, Moreno RD. SNAREs in mammalian sperm: possible implications for fertilization. Dev Biol. 2000; 223(1):54-69. (Clone-specific: Immunofluorescence, Western blot). 查看参考
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Scheller RH. Membrane trafficking in the presynaptic nerve terminal. Neuron. 1995; 14(5):893-897. (Biology). 查看参考
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