The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use at a concentration of 1 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 82 kDa (predicted molecular weight: 82 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
SV2s (Synaptic Vesicle protein 2) are integral membrane glycoproteins present in all synaptic vesicles. They have 12 transmembrane domains predicted by sequence analysis. There are three characterized isoforms, SV2A, SV2B and SV2C. SV2A is expressed ubiquitously throughout the brain. SV2B has a more restricted distribution with varying degrees of coexpression with SV2A. SV2C is more closely related to SV2A but shows a very restricted expression pattern; the highest expression levels being observed in phylogenetically old brain areas like pallidum, the midbrain and the olfactory bulb.
ICC/IF image of ab33892 stained PC12 cells. The cells were 4% PFA fixed (10 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 (ab33892, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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).
Immunohistochemistry (PFA perfusion fixed frozen sections) - SV2C antibody (ab33892)This image is courtesy of an Abreview submitted by Dr Sophie Pezet
ab33892 staining SV2C in mouse brain tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue from 4% PFA perfused animals underwent overnight fixation in 4% paraformaldehyde, cryoprotected in 30% sucrose and cut using cryostat.The primary antibody was diluted, 1/1000 (PBS + 0.3% Triton X100) and incubated with sample for 18 hours at 20°C. An abcam antibody ab60314, Chromeo488 conjugated goat polyclonal to rabbit IgG was used undiluted as secondary. The picture was taken at level of hippocampus.
Couesnon A et al. Preferential entry of botulinum neurotoxin A Hc domain through intestinal crypt cells and targeting to cholinergic neurons of the mouse intestine. PLoS Pathog8:e1002583 (2012).
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