Anti-SNAP25 antibody [4A3] (ab66066)
Key features and details
- Mouse monoclonal [4A3] to SNAP25
- Suitable for: WB, ICC/IF, Flow Cyt
- Reacts with: Mouse, Human, Zebrafish
- Isotype: IgG1
Overview
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Product name
Anti-SNAP25 antibody [4A3]
See all SNAP25 primary antibodies -
Description
Mouse monoclonal [4A3] to SNAP25 -
Host species
Mouse -
Tested applications
Suitable for: WB, ICC/IF, Flow Cytmore details -
Species reactivity
Reacts with: Mouse, Human, Zebrafish
Predicted to work with: Rat, Chicken, Dog, Xenopus laevis, Cynomolgus monkey, Opossum, Orangutan -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: SNAP25 transfected 293T cell lysate; Zebrafish brain homogenate; SH-SY5Y; Mouse brain homogenate. IHC-P: SKNSH cells. Flow cyt: SH-SY5Y cells.
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General notes
This product was changed from ascites to tissue culture supernatant on 4th April 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Constituent: 100% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
4A3 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab66066 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 |
Use at an assay dependent concentration. Detects a band of approximately 27 kDa (predicted molecular weight: 23 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt |
Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
Use at an assay dependent concentration. Detects a band of approximately 27 kDa (predicted molecular weight: 23 kDa). |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt
Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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: 396444 Chicken
- Entrez Gene: 6616 Human
- Entrez Gene: 20614 Mouse
- Entrez Gene: 25012 Rat
- Entrez Gene: 30712 Zebrafish
- Omim: 600322 Human
- SwissProt: P60878 Chicken
- SwissProt: P60880 Human
see all -
Alternative names
- bA416N4.2 antibody
- Bdr antibody
- CMS18 antibody
see all
Images
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All lanes : Anti-SNAP25 antibody [4A3] (ab66066) at 1 µg/ml
Lane 1 : SNAP25 transfected 293T cell lysate
Lane 2 : Non-transfected 293T cell lysate
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG (H&L)-HRP at 1/2500 dilution
Predicted band size: 23 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?This image was generated using the ascites version of the product.
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All lanes : Anti-SNAP25 antibody [4A3] (ab66066) 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 Mouse 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 minutesThis image was generated using the ascites version of the product.
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Overlay histogram showing SH-SY5Y cells stained with ab66066 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab66066, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This image was generated using the ascites version of the product.
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ab66066 stained SKNSH cells. The cells were 4% formaldehyde 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 ab66066 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) 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) at a concentration of 1.43µM.
This image was generated using the ascites version of the product.
Protocols
Datasheets and documents
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Datasheet download
References (8)
ab66066 has been referenced in 8 publications.
- Ponterio G et al. Alpha-Synuclein is Involved in DYT1 Dystonia Striatal Synaptic Dysfunction. Mov Disord 37:949-961 (2022). PubMed: 35420219
- Prikas E et al. Tau target identification reveals NSF-dependent effects on AMPA receptor trafficking and memory formation. EMBO J 41:e10242 (2022). PubMed: 35993331
- Mo J et al. PRRT2 deficiency induces paroxysmal kinesigenic dyskinesia by influencing synaptic function in the primary motor cortex of rats. Neurobiol Dis 121:274-285 (2019). PubMed: 30347267
- Joussain C et al. Botulinum Neurotoxin Light Chains Expressed by Defective Herpes Simplex Virus Type-1 Vectors Cleave SNARE Proteins and Inhibit CGRP Release in Rat Sensory Neurons. Toxins (Basel) 11:N/A (2019). PubMed: 30791373
- Peterson AR & Binder DK Regulation of Synaptosomal GLT-1 and GLAST during Epileptogenesis. Neuroscience 411:185-201 (2019). PubMed: 31158434
- Moscoso-Castro M et al. Cognitive impairments associated with alterations in synaptic proteins induced by the genetic loss of adenosine A2A receptors in mice. Neuropharmacology 126:48-57 (2017). PubMed: 28844595
- Yang H et al. Brain-Specific SNAP-25 Deletion Leads to Elevated Extracellular Glutamate Level and Schizophrenia-Like Behavior in Mice. Neural Plast 2017:4526417 (2017). PubMed: 29318050
- Morató X et al. The Parkinson's disease-associated GPR37 receptor interacts with striatal adenosine A2Areceptor controlling its cell surface expression and function in vivo. Sci Rep 7:9452 (2017). PubMed: 28842709