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  1. Link

    snap25-antibody-epr3275-ab109105.pdf

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Neuroscience Neurotransmission Secretory Vesicles SNAPs & SNAREs
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SNAP25 antibody [EPR3275] (ab109105)

  • Datasheet
  • SDS
Reviews (3)Q&A (1)References (11)

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Flow Cytometry - Anti-SNAP25 antibody [EPR3275] (ab109105)
  • Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105)
  • Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105)
  • Immunocytochemistry/ Immunofluorescence - Anti-SNAP25 antibody [EPR3275] (ab109105)
  • Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105)
  • Anti-SNAP25 antibody [EPR3275] (ab109105)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3275] to SNAP25
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-SNAP25 antibody [EPR3275]
    See all SNAP25 primary antibodies
  • Description

    Rabbit monoclonal [EPR3275] to SNAP25
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human brain, Neuro 2a, mouse brain, mouse hippocampus, rat brain, rat hippocampus cell lysates. ICC/IF: Neuro-2a(Mouse neuroblastoma neuroblast)
  • General notes

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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
    For more information see here.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3275
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmission
    • Secretory Vesicles
    • SNAPs & SNAREs
    • Neuroscience
    • Neurotransmission
    • Secretory Vesicles
    • Other
    • Signal Transduction
    • Protein Trafficking
    • Vesicle Transport
    • SNAPs & SNAREs
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Synapse marker
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Integration of energy metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy
    • Metabolism
    • Types of disease
    • Cancer
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-SNAP25 antibody [EPR3275] - BSA and Azide free (ab171355)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human SNAP25 protein (ab74529)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109105 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
Flow Cyt (Intra)
1/200.
WB (3)
1/1000. Predicted molecular weight: 23 kDa.
ICC/IF
1/250.
Notes
Flow Cyt (Intra)
1/200.
WB
1/1000. Predicted molecular weight: 23 kDa.
ICC/IF
1/250.

Target

  • 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
    modifications

    Palmitoylated. 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.
  • Target information above from: UniProt accession P60880 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Unigene: 45953 Mouse
    • Unigene: 107689 Rat
    see all
  • Alternative names

    • bA416N4.2 antibody
    • Bdr antibody
    • CMS18 antibody
    • dJ1068F16.2 antibody
    • FLJ23079 antibody
    • HGNC:11132 antibody
    • MGC105414 antibody
    • MGC139754 antibody
    • Resistance to inhibitors of cholinesterase 4 homolog antibody
    • RIC 4 antibody
    • RIC-4 antibody
    • RIC4 antibody
    • SEC 9 antibody
    • SEC9 antibody
    • SNAP 25 antibody
    • SNAP antibody
    • SNAP-25 antibody
    • SNAP-25B antibody
    • SNAP25 antibody
    • SNP 25 antibody
    • SNP25 antibody
    • SNP25_HUMAN antibody
    • sp antibody
    • SUP antibody
    • Super protein antibody
    • Synaptosomal associated 25 kDa protein antibody
    • Synaptosomal associated protein antibody
    • Synaptosomal associated protein 25 antibody
    • Synaptosomal associated protein 25kDa antibody
    • Synaptosomal-associated 25 kDa protein antibody
    • Synaptosomal-associated protein 25 antibody
    • Synaptosomal-associated protein antibody
    • Synaptosomal-associated protein, 25-KD antibody
    see all

Images

  • Flow Cytometry - Anti-SNAP25 antibody [EPR3275] (ab109105)
    Flow Cytometry - Anti-SNAP25 antibody [EPR3275] (ab109105)

    Flow Cytometry analysis of Neuro-2a(Mouse neuroblastoma neuroblast) cells labelling SNAP25 with Purified ab109105 at 1:200 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor™ 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105)
    Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105)
    All lanes : Anti-SNAP25 antibody [EPR3275] (ab109105) at 1/1000 dilution (Purified)

    Lane 1 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate
    Lane 2 : Mouse brain lysate
    Lane 3 : Mouse hippocampus lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat hippocampus lysate

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 23 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105)
    Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105)
    Anti-SNAP25 antibody [EPR3275] (ab109105) at 1/1000 dilution (Purified) + Human brain lysate

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 23 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence - Anti-SNAP25 antibody [EPR3275] (ab109105)
    Immunocytochemistry/ Immunofluorescence - Anti-SNAP25 antibody [EPR3275] (ab109105)
    Immunocytochemistry analysis of Neuro-2a(Mouse neuroblastoma neuroblast) cells labeling SNAP25 with Purified ab109105 at 1/250 dilution (8.9 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105)
    Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105)
    All lanes : Anti-SNAP25 antibody [EPR3275] (ab109105) 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 [EPR3275] 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, ab109105 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.

  • Anti-SNAP25 antibody [EPR3275] (ab109105)
    Anti-SNAP25 antibody [EPR3275] (ab109105)

Protocols

  • Flow cytometry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (11)

Publishing research using ab109105? Please let us know so that we can cite the reference in this datasheet.

ab109105 has been referenced in 11 publications.

  • Li Y  et al. Combination of AAV-mediated NUPR1 knockdown and trifluoperazine induces premature senescence in human lung adenocarcinoma A549 cells in nude mice. Oncol Rep 43:681-688 (2020). PubMed: 31922247
  • Wang Y  et al. 27-Hydroxycholesterol Alters Synaptic Structural and Functional Plasticity in Hippocampal Neuronal Cultures. J Neuropathol Exp Neurol 78:238-247 (2019). PubMed: 30753597
  • Cui Z  et al. Nanodiamond autophagy inhibitor allosterically improves the arsenical-based therapy of solid tumors. Nat Commun 9:4347 (2018). PubMed: 30341298
  • Lizarbe B  et al. Neurochemical Modifications in the Hippocampus, Cortex and Hypothalamus of Mice Exposed to Long-Term High-Fat Diet. Front Neurosci 12:985 (2018). PubMed: 30670942
  • Mendsaikhan A  et al. Differences in Gene Expression Profiles and Phenotypes of Differentiated SH-SY5Y Neurons Stably Overexpressing Mitochondrial Ferritin. Front Mol Neurosci 11:470 (2018). PubMed: 30670947
View all Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Western blot abreview for Anti-SNAP25 antibody [EPR3275]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (SHSY5Y cells)
Gel Running Conditions
Reduced Denaturing (4-20% tris-glycine gel)
Loading amount
40 µg
Specification
SHSY5Y cells
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 5%
Read More

Abcam user community

Verified customer

Submitted Dec 30 2020

Western blot abreview for Anti-SNAP25 antibody [EPR3275]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Balb/C mice brain)
Gel Running Conditions
Reduced Denaturing (4-20% tris-glycin gel)
Loading amount
40 µg
Specification
Balb/C mice brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted Dec 29 2020

Western blot abreview for Anti-SNAP25 antibody [EPR3275]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Purified protein (Synaptosomes)
Gel Running Conditions
Reduced Non-Denaturing (Native) (4–20% Mini-PROTEAN TGX Precast Protein Gel)
Loading amount
10 µg
Specification
Synaptosomes
Blocking step
Odyssey Blocking Buffer (PBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 23°C
Read More

Abcam user community

Verified customer

Submitted Apr 15 2019

Question

Hi, I am wondering if you could provide us with the information on the epitope sequence/region of SNAP-25 that this antibody recognizes (eg. N-terminus, C-terminus..).

Read More

Abcam community

Verified customer

Asked on Jul 06 2012

Answer

Thank you for contacting us.

The peptide maps between amino acids 180 - 206 of human SNAP-25, at the C-terminus (UniProt: P60880).

Please let me know if you have any questions.


We hope this response has sufficiently answered your questions.

Read More

Abcam Scientific Support

Answered on Jul 06 2012

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