The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesICC/IF: Use at an assay dependent dilution (PMID 19448629).
IHC-Fr: Use at an assay dependent dilution (PMID 20010821).
WB: Use at a concentration of 1 - 5 µg/ml. Predicted molecular weight: 48 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
RelevanceSynaptotagmin 4 (SYT4) belongs to the synaptotagmin family of multi-domained, integral membrane proteins of synaptic vesicles. It may be involved in Ca(2+)-dependent exocytosis of secretory vesicles through Ca(2+) and phospholipid binding to the C2 domain or may serve as Ca(2+) sensors in the process of vesicular trafficking and exocytosis. It is primarily expressed in the brain.
ICC/IF image of ab57473 stained SHSY5Y cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab57473, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-Synaptotagmin 4 antibody (ab57473)
This product has been referenced in:
Du W et al. HID-1 is required for homotypic fusion of immature secretory granules during maturation. Elife5:N/A (2016).
Read more (PubMed: 27751232) »
Johnson SL et al. Synaptotagmin IV determines the linear Ca2+ dependence of vesicle fusion at auditory ribbon synapses. Nat Neurosci13:45-52 (2010).
Read more (PubMed: 20010821) »