Recombinant Anti-SNAP25 antibody [EP3274] (ab108990)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP3274] to SNAP25
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SNAP25 antibody [EP3274]
See all SNAP25 primary antibodies -
Description
Rabbit monoclonal [EP3274] to SNAP25 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Fetal brain, HeLa, SH-SY5Y, U87-MG, fetal hippocampus, fetal cerebellum, and Neuro-2a cell lysates; Human brain tissue; Human cerebellum tissue, mouse and rat brain tissue lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP3274 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab108990 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 |
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WB |
1/1000 - 1/10000. Detects a band of approximately 25 kDa (predicted molecular weight: 23 kDa).
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IHC-P | (2) |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Antigen retrieval is recommended. |
Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 25 kDa (predicted molecular weight: 23 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Antigen retrieval is recommended. |
Target
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Function
t-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 specificity
Neurons of the neocortex, hippocampus, piriform cortex, anterior thalamic nuclei, pontine nuclei, and granule cells of the cerebellum. -
Sequence similarities
Belongs 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 localization
Cytoplasm > 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: 6616 Human
- Entrez Gene: 20614 Mouse
- Entrez Gene: 25012 Rat
- Omim: 600322 Human
- SwissProt: P60880 Human
- SwissProt: P60879 Mouse
- SwissProt: P60881 Rat
- Unigene: 167317 Human
see all -
Alternative names
- bA416N4.2 antibody
- Bdr antibody
- CMS18 antibody
see all
Images
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All lanes : Anti-SNAP25 antibody [EP3274] (ab108990) at 1/1000 dilution
Lane 1 : Wild-type SH-SY5Y cell lysate
Lane 2 : SNAP25 knockout SH-SY5Y cell lysate
Lane 3 : Neuro-2a cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-SNAP25 antibody [EP3274] staining at 1/1000 dilution, shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, ab108990 was shown to bind specifically to SNAP25. A band was observed at 27 kDa in wild-type SH-SY5Y cell lysates with no signal observed at this size in SNAP25 knockout cell line ab280041 (knockout cell lysate ab280100). To generate this image, wild-type and SNAP25 knockout SH-SY5Y cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-SNAP25 antibody [EP3274] (ab108990) at 1/1000 dilution
Lane 1 : Fetal brain lysate
Lane 2 : HeLa cell lysate
Lane 3 : SH-SY5Y cell lysate
Lane 4 : U87-MG cell lysate
Lane 5 : Fetal hippocampus lysate
Lane 6 : Fetal cerebellum lysate
Lane 7 : Neuro-2a cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 23 kDa -
ab108990, at 1/100 dilution, staining SNAP25 in paraffin-embedded Human brain tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-SNAP25 antibody [EP3274] (ab108990) at 1/2000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Mouse kidney tissue lysate
Lane 4 : Mouse spleen tissue lysate
Lane 5 : Rat brain tissue lysate
Lane 6 : Rat heart tissue lysate
Lane 7 : Rat kidney tissue lysate
Lane 8 : Rat spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration: 5% NFDM /TBST
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ab108990, at 1/100 dilution, staining SNAP25 in paraffin-embedded Human cerebellum tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab108990 showing negative staining in human Normal colon tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab108990 showing negative staining in human Normal spleen tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab108990 showing negative staining in human Skeletal muscle tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (9)
ab108990 has been referenced in 9 publications.
- Sadeghi L et al. Proteomic profiling of the rat hippocampus from the kindling and pilocarpine models of epilepsy: potential targets in calcium regulatory network. Sci Rep 11:8252 (2021). PubMed: 33859251
- Lee SH et al. APP Family Regulates Neuronal Excitability and Synaptic Plasticity but Not Neuronal Survival. Neuron 108:676-690.e8 (2020). PubMed: 32891188
- Liao Y et al. d-Glucuronolactone attenuates para-xylene-induced defects in neuronal development and plasticity in Xenopus tectum in vivo. Toxicology 430:152341 (2020). PubMed: 31811891
- Kang J & Shen J Cell-autonomous role of Presenilin in age-dependent survival of cortical interneurons. Mol Neurodegener 15:72 (2020). PubMed: 33302995
- Shi TY et al. Kainate receptor mediated presynaptic LTP in agranular insular cortex contributes to fear and anxiety in mice. Neuropharmacology 128:388-400 (2018). WB . PubMed: 29104074
- Almandoz-Gil L et al. In Situ Proximity Ligation Assay Reveals Co-Localization of Alpha-Synuclein and SNARE Proteins in Murine Primary Neurons. Front Neurol 9:180 (2018). Mouse . PubMed: 29623065
- Geerts CJ et al. Tomosyn associates with secretory vesicles in neurons through its N- and C-terminal domains. PLoS One 12:e0180912 (2017). PubMed: 28746398
- Jin WS et al. Peritoneal dialysis reduces amyloid-beta plasma levels in humans and attenuates Alzheimer-associated phenotypes in an APP/PS1 mouse model. Acta Neuropathol 134:207-220 (2017). PubMed: 28477083
- Lue LF et al. TREM2 Protein Expression Changes Correlate with Alzheimer's Disease Neurodegenerative Pathologies in Post-Mortem Temporal Cortices. Brain Pathol 25:469-80 (2015). IHC-FrFl ; Human . PubMed: 25186950