Immunofluorescence analysis of VAMP was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with VAMP / synaptobrevin (SP10) Mouse Monoclonal Antibody (P21941) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Hamster, Human, Mouse, Pig, Rabbit, Rat|
|Host / Isotype||Mouse / IgM|
|Immunogen||Synaptic immunoprecipitate (crude) from human brain.|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||1:500-1:2000|
* 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.
Synaptophysin is a 38kDa glycoprotein present in the membrane of neuronal presynaptic vesicles in brain, spinal cord, retina, vesicles of adrenal medulla, neuromuscular junctions, and endocrine cells. It is also expressed by neuroendocrine cells throughout the body, both normal and neoplastic. Synaptophysin is a useful marker for the identification of normal neuroendocrine cells and neuroendocrine neoplasms. This antibody reacts with neuroendocrine cells of human adrenal medulla, carotid body, skin, pituitary gland, thyroid, lung, pancreas, and gastointestinal mucosa. It also reacts with a wide spectrum of neuroendocrine neoplasms of neural type including neuroblastomas, ganglioneuroblastomas, ganglioneuromas, pheochromocytomas, chromaffin, and non-chromaffin paragangliomas.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.