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S9684

Sigma-Aldrich

Anti-SNAP-25 antibody produced in rabbit

IgG fraction of antiserum, buffered aqueous solution

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About This Item

MDL number:
UNSPSC Code:
12352203
NACRES:
NA.46

biological source

rabbit

Quality Level

conjugate

unconjugated

antibody form

IgG fraction of antiserum

antibody product type

primary antibodies

clone

polyclonal

form

buffered aqueous solution

mol wt

antigen 25 kDa

species reactivity

rat, mouse

packaging

antibody small pack of 25 μL

technique(s)

immunohistochemistry (formalin-fixed, paraffin-embedded sections): 1:5,000 using rat cerebellum sections
microarray: suitable
western blot: 1:10,000 using rat pheochromocytoma PC12 cell extract
western blot: 1:5,000 using mouse brain extract

UniProt accession no.

shipped in

dry ice

storage temp.

−20°C

target post-translational modification

unmodified

Gene Information

human ... SNAP25(6616)
mouse ... Snap25(20614)
rat ... Snap25(25012)

General description

SNAP-25 (Synaptosome-associated protein of 25 kDa) is a membrane bound, presynaptic nerve terminal protein, that plays an essential role in synaptic vesicle fusion and exocytosis. The gene encoding SNAP-25 is mapped to human chromosome 20.

Anti-SNAP-25 is developed in rabbit using a synthetic peptide corresponding to the N-terminus of human SNAP-25 conjugated to KLH as immunogen. This sequence is identical in SNAP-25 alternatively spliced forms SNAP-25A and SNAP-25B, in mouse, rat and chicken SNAP-25 and highly conserved (80-85%) in goldfish and zebrafish SNAP25. Whole antiserum is fractionated and then further purified by ion-exchange chromatography to provide the IgG fraction of antiserum that is essentially free of other rabbit serum proteins. Anti-SNAP-25 recognizes mouse SNAP-25 (25 kD). The antibody cross-reacts with rat SNAP-25. Applications include the detection and localization of SNAP-25 (25 kDa) by immunoblotting and immunohistochemistry. Staining of SNAP-25 in immunoblotting is specifically inhibited with SNAP-25 immunizing peptide (SNAP-25, mouse)

Specificity

The immunogen sequence is identical in SNAP-25A and SNAP-25B splice variants in mouse, rat and chicken SNAP-25, and is highly conserved (70-80%) in bovine, goldfish, torpedo and Drosophila SNAP-25.

Immunogen

Synthetic peptide corresponding to the N-terminal of human SNAP-25 (synaptosome-associated protein-25) amino acids conjugated to KLH.

Application

Anti-SNAP-25 antibody produced in rabbit has been used in:
  • the primary detection of SNAP-25
  • enzyme-linked immunosorbent assay in human neuroblastoma cell lines
  • immunohistochemistry
  • in human neuronal cells using western blotting

Biochem/physiol Actions

Synaptosome-associated protein of 25 kDa codes for a protein impacting axonal growth, synaptic plasticity, and presynaptic neurons necessary for the regulation of neurotransmitter release.

The molecular events leading to neurotransmitter release in the synaptic cleft are complex, involving multiple interacting proteins, generically termed SNAP receptors (SNAREs). It has been suggested that SNAP-25 and syntaxin on the neuronal plasma membrane (t-SNARE) and synaptobrevin/VAMP on the synaptic vesicle (v-SNARE) form a stable ternary complex. This core complex serves as a docking complex for two additional membrane fusion proteins, β-SNAP and NSF. ATP hydrolysis by NSF causes dissociation of the complex during priming of the exocytosis machinery. SNAP-25 induced reassembly and interaction with synaptotagmin (Syt), is thought to drive the Ca2+ -triggered vesicle-plasma membrane fusion and exocytosis. SNAP-25 has a key role in both developing and mature neurons. During development, SNAP-25 expression correlates with synaptogenesis, axonal growth and neuronal maturation and is found mainly in cell bodies of neonatal brain. In the adult nervous system, SNAP-25 is localized to presynaptic nerve terminals where it is conveyed by fast axonal transport.

SNAP-25 consists of two alternatively spliced isoforms SNAP-25a and SNAP-25b, differentially expressed in neurons and neuroendocrine cells. SNAP-25a and SNAP-25b differ by nine amino acids in the central domain. Two of these residues alter the relative positioning of clustered cysteine residues that are required for post-translational palmitoylation implicated in membrane anchoring, suggesting that the two SNAP-25 isoforms may play distinct roles in vesicular fusion events.

Physical form

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.

Storage and Stability

For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing is not recommended. Storage in "frost-free" freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class Code

12 - Non Combustible Liquids

WGK

nwg

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Synaptotagmin interaction with SNAP-25 governs vesicle docking, priming, and fusion triggering.
Mohrmann R, et al.
The Journal of Neuroscience, 33(36), 14417-14430 (2013)
Membrane trafficking in neuronal development: ins and outs of neural connectivity.
Winkle CC and Gupton SL
International Review of Cell and Molecular Biology, 322, 247-280 (2016)
The 25-kDa synaptosome-associated protein (SNAP-25) binds and inhibits delayed rectifier potassium channels in secretory cells.
Ji J, et al.
The Journal of Biological Chemistry, 277(23), 20195-20204 (2002)
Cleavage of neuronal synaptosomal-associated protein of 25 kDa by exogenous matrix metalloproteinase-7.
Szklarczyk A , et al.
Journal of Neurochemistry, 102(4), 1256-1263 (2007)
Association of syntaxin with SNAP-25 and VAMP (synaptobrevin) during axonal transport.
Shiff G and Morel N
Journal of Neuroscience Research, 48(4), 313-323 (1997)

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