SYT7 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Synaptotagmin VII (SYT7) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Synaptotagmin VII (SYT7) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Ca (2+) sensor involved in Ca (2+) -dependent exocytosis of secretory and synaptic vesicles through Ca (2+) and phospholipid binding to the C2 domain (By similarity) . Ca (2+) induces binding of the C2-domains to phospholipid membranes and to assembled SNARE-complexes; both actions contribute to triggering exocytosis (By similarity) . SYT7 binds Ca (2+) with high affinity and slow kinetics compared to other synaptotagmins (By similarity) . Involved in Ca (2+) -triggered lysosomal exocytosis, a major component of the plasma membrane repair (PubMed:11342594) . Ca (2+) -regulated delivery of lysosomal membranes to the cell surface is also involved in the phagocytic uptake of particles by macrophages (By similarity) . Ca (2+) -triggered lysosomal exocytosis also plays a role in bone remodeling by regulating secretory pathways in osteoclasts and osteoblasts (By similarity) . In case of infection, involved in participates cell invasion by Trypanosoma cruzi via Ca (2+) -triggered lysosomal exocytosis (PubMed:11342594, PubMed:15811535) . Involved in cholesterol transport from lysosome to peroxisome by promoting membrane contacts between lysosomes and peroxisomes: probably acts by promoting vesicle fusion by binding phosphatidylinositol-4,5-bisphosphate on peroxisomal membranes (By similarity) . Acts as a key mediator of synaptic facilitation, a process also named short-term synaptic potentiation: synaptic facilitation takes place at synapses with a low initial release probability and is caused by influx of Ca (2+) into the axon terminal after spike generation, increasing the release probability of neurotransmitters (By similarity) . Probably mediates synaptic facilitation by directly increasing the probability of release (By similarity) . May also contribute to synaptic facilitation by regulating synaptic vesicle replenishment, a process required to ensure that synaptic vesicles are ready for the arrival of the next action potential: SYT7 is required for synaptic vesicle replenishment by acting as a sensor for Ca (2+) and by forming a complex with calmodulin (By similarity) . Also acts as a regulator of Ca (2+) -dependent insulin and glucagon secretion in beta-cells (By similarity) . Triggers exocytosis by promoting fusion pore opening and fusion pore expansion in chromaffin cells (By similarity) . Also regulates the secretion of some non-synaptic secretory granules of specialized cells (By similarity) .

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Synaptotagmin VII (SYT7) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Synaptotagmin VII (SYT7) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Synaptotagmin VII (SYT7), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Synaptotagmin VII (SYT7) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Component Amount Anti-SYT7 Microplate 96 Wells (12 x 8 Well Strips) SYT7 Lyophilized Standard 2 x 20 ng 100X Biotinylated SYT7 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-SYT7 Microplate 96 Wells (12 x 8 Well Strips) Anti-SYT7 Microplate 96 Wells (12 x 8 Well Strips) SYT7 Lyophilized Standard 2 x 20 ng SYT7 Lyophilized Standard 2 x 20 ng 100X Biotinylated SYT7 Detector Antibody 120 uL 100X Biotinylated SYT7 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Synaptotagmin VII (SYT7) . No significant cross-reactivity or interference between Synaptotagmin VII (SYT7) and analogues was observed.