Anti-SNAP25 antibody (ab5666)
- Product nameAnti-SNAP25 antibodySee all SNAP25 primary antibodies ...
- DescriptionRabbit polyclonal to SNAP25
- Tested applicationsIP, ICC, WB more details
- Species reactivityReacts with: Mouse, Rat, Cow
Synthetic peptide: ANQRATKMLGSG, corresponding to amino acids 195-206 of Mouse SNAP25. This sequence is completely conserved between mouse, rat, human, chicken, and goldfish.
- Positive controlICC/IF: PC12 cell line.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.05% Sodium Azide
Constituents: PBS, 1mg/ml BSA
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Integration of energy
Our Abpromise guarantee covers the use of ab5666 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||IP: Use a concentration of 5 µg/ml.|
|ICC||ICC: 1/100. Immunofluorescence staining of SNAP-25 in cultured rat hippocampal cells with ab5666 yields a pattern consistent with cytoplasmic and plasma membrane staining.|
|WB||WB: 1/2000. Detects a band of approximately 25 kDa.Can be blocked with SNAP25 peptide (ab5843). representing SNAP-25 whole rat brain extract.|
- Functiont-SNARE involved in the molecular regulation of neurotransmitter release. May play an important role in the synaptic function of specific neuronal systems. Associates with proteins involved in vesicle docking and membrane fusion. Regulates plasma membrane recycling through its interaction with CENPF.
- Tissue specificityNeurons of the neocortex, hippocampus, piriform cortex, anterior thalamic nuclei, pontine nuclei, and granule cells of the cerebellum.
- Sequence similaritiesBelongs to the SNAP-25 family.
Contains 2 t-SNARE coiled-coil homology domains.
modificationsPalmitoylated. Cys-85 appears to be the main site, and palmitoylation is required for membrane association.
- Cellular localizationCytoplasm > perinuclear region. Cell membrane. Cell junction > synapse > synaptosome. Membrane association requires palmitoylation. Expressed throughout cytoplasm, concentrating at the perinuclear region.
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Anti-SNAP25 antibody images
Immunlocalization of SNAP-25 in cultured rat hippocampal cells using ab5666.
Anti-SNAP25 antibody (ab5666) at 1/10000 dilution + Whole tissue lysate prepared from mouse hippocampus.
HRP conjugated goat anti-rabbit Ig. at 1/10000 dilution
developed using the ECL technique
Performed under reducing conditions.
Observed band size : 25 kDa (why is the actual band size different from the predicted?)
Exposure time : 20 seconds
This image is courtesy of an anonymous Abreview
ICC/IF image of ab5666 stained PC12 cells. The cells were 4% paraformaldehyde 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 (ab5666, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
SNAP25 was immunoprecipitated using 0.5mg Mouse Brain tissue lysate, 5µg of Rabbit polyclonal to SNAP25 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab5666.
Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
Band: 25kDa; SNAP25
References for Anti-SNAP25 antibody (ab5666)
This product has been referenced in:
- Tozawa T et al. The shortest isoform of dystrophin (Dp40) interacts with a group of presynaptic proteins to form a presumptive novel complex in the mouse brain. Mol Neurobiol 45:287-97 (2012). WB, ICC/IF ; Mouse . Read more (PubMed: 22258561) »
- Ramos C et al. Native pre-synaptic metabotropic glutamate receptor 4 (mGluR4)interacts with exocytosis proteins in rat cerebellum. J Biol Chem : (2012). WB . Read more (PubMed: 22528491) »