Anti-SNAP25 antibody (ab41455)
- Datasheet
- References (9)
- Protocols
Overview
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Product nameAnti-SNAP25 antibody
See all SNAP25 primary antibodies -
DescriptionRabbit polyclonal to SNAP25
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Host speciesRabbit
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Tested applicationsSuitable for: IP, IHC-FoFr, WB, ICC/IFmore details
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Species reactivityReacts with: Mouse, Rat, Human, Zebrafish
Predicted to work with: Chicken, Cow
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human SNAP25.
Read Abcam's proprietary immunogen policy (Peptide available as ab41724.) -
Positive control
- Recombinant Human SNAP25 protein (ab51048) can be used as a positive control in WB. This antibody gave a positive signal in the following lysates: Spinal cord (Rat) tissue lysate and Hippocampus (Mouse) tissue Lysate. ICC/IF: PC12 cell line.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab41455 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IP | Use a concentration of 5 µg/ml. | |
| IHC-FoFr | 1/1000. | |
| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 23 kDa). | |
| ICC/IF | Use a concentration of 5 µg/ml. |
Target
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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.
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Tissue specificityNeurons of the neocortex, hippocampus, piriform cortex, anterior thalamic nuclei, pontine nuclei, and granule cells of the cerebellum.
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Sequence similaritiesBelongs to the SNAP-25 family.
Contains 2 t-SNARE coiled-coil homology domains. -
Post-translational
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.
- Information by UniProt
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Database links
- Entrez Gene: 396444 Chicken
- Entrez Gene: 540853 Cow
- Entrez Gene: 6616 Human
- Entrez Gene: 20614 Mouse
- Entrez Gene: 25012 Rat
- Entrez Gene: 30712 Zebrafish
- Omim: 600322 Human
- SwissProt: P60878 Chicken
see all -
Alternative names
- bA416N4.2 antibody
- Bdr antibody
- CMS18 antibody
see all
Images
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Western blot - Anti-SNAP25 antibody (ab41455)All lanes : Anti-SNAP25 antibody (ab41455) at 1 µg/ml
Lane 1 : Spinal cord (Rat) tissue lysate
Lane 2 : Hippocampus (Mouse) tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted? -
Western blot - Anti-SNAP25 antibody (ab41455) -
Immunocytochemistry/ Immunofluorescence - Anti-SNAP25 antibody (ab41455)ab4455 staining SNAP25 in PC12 cells. The cells were fixed with 100% methanol (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 ab41455 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
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Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-SNAP25 antibody (ab41455)This image is courtesy of an Abreview submitted by Dr Sophie Pezetab41455 staining SNAP25 in rat 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, 1/3000 (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, diluted 1/1000 was used as secondary.
The picture shows the staining obtained at the level of the hippocampus. The staining was very strong, located in the whole brain.
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Western blot - Anti-SNAP25 antibody (ab41455)All lanes : Anti-SNAP25 antibody (ab41455) at 1 µg/ml
Lane 1 : Marker
Lane 2 : Zebrafish brain homogenate at 20 µg
Lane 3 : SH-SY5Y (Human neuroblastoma cell line) whole cell lysate at 20 µg
Lane 4 : Mouse brain homogenate at 20 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes -
Immunoprecipitation - Anti-SNAP25 antibody (ab41455)SNAP25 was immunoprecipitated using 0.5mg Rat Spinal Cord 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, Rat Spinal Cord 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 ab41455.
Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
Band: 26kDa; SNAP25
Protocols
Datasheets and documents
References
This product has been referenced in:
- Pan Y et al. Pancreatic fibroblast growth factor 21 protects against type 2 diabetes in mice by promoting insulin expression and secretion in a PI3K/Akt signaling-dependent manner. J Cell Mol Med N/A:N/A (2018). Read more (PubMed: 30461198) »
- Shirafuji T et al. The Role of Cysteine String Protein a Phosphorylation at Serine 10 and 34 by Protein Kinase C? for Presynaptic Maintenance. J Neurosci 38:278-290 (2018). Read more (PubMed: 29167402) »
