Technical Data
S9108-98
Synaptotagmin (Syt)
Description:
Synaptotagmins (Syt) are membrane-trafficking proteins characterized by an N-terminal transmembrane region, a variable linker and two C-terminal C2-domains, C2A and C2B. At least twelve synaptotagamins are expressed in vertebrates and can be found in both vesicular and plasma membranes. Synaptotagamins are membrane-bound Ca2+-sensor proteins that bind Ca2+ with low affinity. Although all Syts bind Ca2+ with the same cooperativity and participate in Ca2+ dependent phospholipid binding they have distinct Ca2+-binding properties and affinities (3). Two broad subfamilies have been proposed based on expression pattern. Syt 1-5, 10, 11 and Srg 1 are primarily expressed in nervous and other endocrine tissues while Syt 69 are ubiquitously expressed (4). The calcium-dependent, lipid-binding properties of Syt 1 suggest its involvement in lipid rearrangements required during synaptic vesicle fusion (3). The three cytosolic loops that form the Ca2+ binding pocket in the C2A domain insert into phospholipid bilayers upon Ca2+ binding. Ca2+ binding also triggers oligimerization of Syt via the C2B domain (4). Plasma membrane Syts have higher Ca2+ affinity than vesicular Syts and are are proposed to be more important for endocrine exocytosis, while vesicular Syts are thought to be more important in fast synaptic exocytosis. The distinct properties, combinantions and relative abundance of various Syts may influence the Ca2+ responses of synapses (3). Syts have also been shown to interact with neural proteins in the absence of calcium, suggesting roles in presynaptic functions. These interactions include binding to calcium channels, sodium channels, the clatherin adapter AP-2, the stoned endocytotic proteins, beta-SNAP, neurexins, calmodulin, inositol polyphosphates, SYNCRIP and polyphosphoinositides (4).

Applications:
Suitable for use in Western Blot, Immunoprecipitation, Immunocytochemistry and Immunohistochemistry.
Other applications not tested.

Recommended Dilution:
Western Blot (Colorimetric): 0.1ug/mL Detects synaptotagmin in ~20ug of rat bain tissue extract.
Immunocytochemistry: 1:500
Optimal dilutions to be determined by the researcher.

Positive Controls:
Mouse Brain Tissue Extract
Rat Brain Tissue Extract

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG2b4E207Affinity Purified
SizeStorageShippingSourceHost
50ug 4C (-20C Glycerol)Blue IceRatMouse
Concentration:
~1mg/ml
Immunogen:
Rat brain synaptic junctional protein complexes
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a liquid in PBS, pH7.2, 0.1mM PMSF, 50% glycerol.
Specificity:
Detects an ~65kD protein, corresponding to the apparent molecular mass of synaptotagmin on SDS-PAGE Immunoblots. Species Crossreactivity: Mouse, rat, bovine, monkey and fish (shark).
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Matthew, W. D., Tsavaler, L., Reichardt, L. F. (1981) J Cell Biol 91: 257-269. 2. Koontz, M.A., et al. (1993) Synapse 14: 268-282. 3. Sudhof, T.C. (2002) J Biol Chem 277(10): 7629-7632. 4. Adolfsen, B., Littleton, J.T. (2001) Cell Mol Life Sci 58: 393-402. 5. Ibata, K., Fukuda, M., Hamada, T., Kabayama, H., Mikoshiba, K. (2000) J Neurochem 74(2): 518-526. 6. Fukuda, M., Kowalchyk, J.A., Zhang, X., Martin, T.F.J., Mikoshiba, K. (2002) J Biol Chem 277(7): 4601-4604.