Western blot analysis of phospho-synapsin 1 was performed by loading 40ug of mouse brain cortex tissue lysates onto a 4-15% gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/PBS at room temperature. Phospho-Synapsin 1 was detected using a phospho-synapsin 1 rabbit polyclonal antibody (Product # PA1-4698) at a dilution of 1;1000 in blocking buffer for 2 hours at room temperature, followed by a HRP conjugated secondary antibody at a dilution of 1:10000 for 1 hour at room temperature. Chemiluminescent detection was performed.
|Tested species reactivity||Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phospho-peptide corresponding to amino acid residues surrounding Ser62/67 of rat SYN1 conjugated to KLH|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is predicted to react with mouse and bovine based on 100% sequence homology.
In Western blot, this antibody is specific for ~78 kDa synapsin I doublet phosphorylated at Ser 62 and 67. Immunolabeling is blocked by the phosphopeptide used as the antigen but not by the corresponding dephosphopeptide.
Synapsin I plays a key role in synaptic plasticity in brain (Feng et al., 2002; Nayak et al., 1996). This effect is due in large part to the ability of the synapsins to regulate the availability of synaptic vesicles for release. The role of synapsin in synaptic plasticity and in synaptogensis is regulated by phosphorylation (Jovanovic et al., 2001; Kao et al., 2002). Ser 549 along with Ser 62 and Ser 67 are the sites of synapsin I that are phosphorylated by MAP kinase (Jovanovic et al., 1996). Phosphorylation and subsequent dephosphorylation of this site is thought to play a key role in synaptic vesicle trafficking.
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